WO2018006063A1

WO2018006063A1 - Compositions and methods for the treatment of bacterial infections

Info

Publication number: WO2018006063A1
Application number: PCT/US2017/040467
Authority: WO
Inventors: James M. Balkovec; Daniel C. BENSEN; Timothy Blizzard; Allen Borchardt; Thomas P. Brady; Zhi-yong CHEN; Quyen-Quyen Thuy Do; Wanlong Jiang; Thanh Lam; Jeffrey B. LOCKE; Alain Noncovich; Leslie W. TARI
Original assignee: Cidara Therapeutics, Inc.
Priority date: 2016-07-01
Filing date: 2017-06-30
Publication date: 2018-01-04

Abstract

Compositions and methods for the treatment of bacterial infections include compounds containing dimers of cyclic heptapeptides conjugated to one or more monosaccharide or oligosaccharide moieties. In particular, compounds can be used in the treatment of bacterial infections caused by Gram-negative bacteria.

Description

COMPOSITIONS AND METHODS FOR THE TREATMENT OF BACTERIAL INFECTIONS

Background

The need for novel antibacterial treatments for bacterial infections is significant and especially critical in the medical field. Antibacterial resistance is a serious global healthcare threat. Polymyxins are a class of antibiotics that exhibit potent antibacterial activities against Gram-negative bacteria. However, the use of polymyxins as an antibiotic has been limited due to the associated toxicity and adverse effects (e.g., nephrotoxicity). Moreover, mcr- 1, a plasmid-borne gene conferring bacterial resistance to polymyxins, has a high potential for dissemination and further threatens the efficacy of this class of antibiotics.

Because of the shortcomings of existing antibacterial treatments, combined with the emergence of multidrug-resistant Gram-negative bacteria, there is a need in the art for improved antibacterial therapies having greater efficacy, bioavailability, and reduced toxicity. Summary

The disclosure relates to compounds, compositions, and methods for inhibiting bacterial growth (e.g., Gram-negative bacterial growth) and for the treatment of bacterial infections (e.g., Gram-negative bacterial infections). In particular, such compounds contain dimers of cyclic heptapeptides conjugated to one or more monosaccharide or oligosaccharide moieties, which interact, directly or indirectly, with an immune cell. In certain embodiments, compounds containing dimers of cyclic heptapeptides may be conjugated to at least 2 (e.g., 2-1 0, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, or 2-3) monosaccharide or oligosaccharide moieties (e.g., rhamnose). In certain embodiments, compounds containing dimers of cyclic

heptapeptides may be conjugated to at least 3 (e.g., 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, or 3-4) monosaccharide or oligosaccharide moieties (e.g., rhamnose). In yet other embodiments, compounds containing dimers of cyclic heptapeptides may be conjugated to at least 4 (e.g., 4-10, 4-9, 4-8, 4-7, 4-6, or 4-5)

monosaccharide or oligosaccharide moieties (e.g., rhamnose). In particular embodiments, compounds containing dimers of cyclic heptapeptides may be conjugated to two monosaccharide or oligosaccharide moieties (e.g., rhamnose). Cyclic heptapeptides bind to lipopolysaccharides (LPS) in the cell membrane of Gram-negative bacteria to disrupt and permeabilize the cell membrane, leading to cell death and/or sensitization to other antibiotics.

In one aspect, the disclosure features a compound described by formula (I):

(E)n

M2 L' M1

(I)

wherein M1 includes a first cyclic heptapeptide including a linking nitrogen and M2 includes a second cyclic heptapeptide including a linking nitrogen; each E is, independently, a monosaccharide or oligosaccharide moiety; L' is a linker covalently attached to E and to the linking nitrogen in each of M1 and M2; and n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

In some embodiments, L' in formula (I) is described by formula (L):

(L)

wherein L is a remainder of L'; A1 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M1 or is absent; and A2 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M2 or is absent.

In some embodiments, the compound is described by formula (II):

M2 A2 A1 M1

(II)

wherein L is a remainder of L'; n is 1 , 2, 3, or 4 (e.g., 1 or 2) ; each of R¹ , R¹², R'¹ , and R'¹² is, independently, a lipophilic moiety, a polar moiety, or H; each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is, independently, optionally substituted C1 -C5 alkamino, a polar moiety, a positively charged moiety, or H; each of R¹⁵ and R'¹⁵ is, independently, a lipophilic moiety or a polar moiety; each of R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R'², R'³, R'⁴, R'⁵, R'⁶, R'⁷, R'⁸, R'⁹, and R'¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20

heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; or two R groups on the same or adjacent atoms join to form an optionally substituted 5-8 membered ring; each of a', b', c', a, b, and c is, independently, 0 or 1 ; each of N¹ , N², N³, N⁴, N'¹ , N'², N'³, and N'⁴ is a nitrogen atom ; each of C¹ , C², C³, C'¹ , C'², and C'³ is a carbon atom, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound includes at least one optionally substituted 5-8 membered ring formed by joining (i) R², R³, and C¹ ; (ii) R³, R⁴, N¹ , and C¹ ; (iii) R⁵, R⁶, and C²; (iv) R⁶, R⁷, N², and C²; (v) R⁸, R⁹, and C³; (vi) R⁹, R¹⁰, N³, and C³; (vii) R'², R'³, and C'¹ ; (viii) R'³, R'⁴, N'¹ , and C'¹ ; (ix) R'⁵, R'⁶, and C'²; (x) R'⁶, R'⁷, N'², and C'²; (xi) R'⁸, R'⁹, and C'³; or (xii) R'⁹, R'¹⁰, N'³, and C'³. 1 some embodiments, the compound is described by formula

(III)

or a pharmaceutically acceptable salt thereof.

In some embodiments, each of R¹ , R¹², R'¹ , and R'¹² is, independently, a lipophilic moiety; each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is, independently, optionally substituted C1 -C5 alkamino, a polar moiety, or a positively charged moiety; and/or each of R¹⁵ and R'¹⁵ is, independently, a polar moiety.

In some embodiments, each of R¹ and R¹² is a lipophilic moiety. In some embodiments, each of R'¹ and R'¹² is a lipophilic moiety. In some embodiments, each lipophilic moiety is, independently, optionally substituted C1 -C20 alkyi, optionally substituted C5-C15 aryl, optionally substituted C6-C35 alkaryl, or optionally C5-C1 0 substituted heteroaryl. In some embodiments, each lipophilic moiety is, independently, C1 -C8 alkyi, methyl substituted C2-C4 alkyi, (C1 -C10)alkylene(C6)aryl, phenyl substituted (C1 -C10)alkylene(C6)aryl, or alkyi substituted C4-C9 heteroaryl. In some embodiments, each lipophilic moiety is, independently, benzyl, isobutyl, sec-butyl, isopropyl, n-propyl, methyl, biphenylmethyl, n-octyl, or methyl substituted indolyl.

In some embodiments, each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is independently optionally substituted C1 -C5 alkamino. In some embodiments, each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is

In some embodiments, each of R¹⁵ and R'¹⁵ is a polar moiety. In some embodiments, each polar moiety includes a hydroxyl group, a carboxylic acid group, an ester group, or an amide group. In some embodiments, each polar moiety is hydroxyl substituted C1 -C4 alkyi. In some embodiments, each polar moiety is CH(CH₃)OH.

In some embodiments, the compound is described by formula (IV):

(IV) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl, or a

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (IV-1 ):

(IV-1 )

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl, or a

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (IV-2):

(IV-2)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl, or a

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (V):

(V) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (V-1 ):

(V-1 )

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; or a

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (V-2):

(V-2)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; or a

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (V-3):

(V-3)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (V-4):

(V-4)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (V-5):

(V-5)

pharmaceutically acceptable salt thereof. In some embodiments, the compound is described by formula (VI):

(VI)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R⁸, R'², R'⁶, and R'⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (VI-1 ):

(VI-1 )

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (VI-2):

(VI-2)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (VI-3):

(VI-3)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (VI-4):

(VI-4)

pharmaceutically acceptable salt thereof. In some embodiments, the compound is described by formula (VI-5):

(VI-5)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (VII):

(VII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R⁸, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 - C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (VIII):

(VIII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R⁸, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (IX):

(IX)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (IX-1 ):

(IX-1 )

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (IX-2):

(IX-2)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (X):

(X)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R² and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XI):

(XI)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R² and R⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XI-1 ):

(XI-1 ) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; or a

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XI-2):

(XI-2)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XII):

(XII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R² and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; R⁶, R⁷, N², and C² together form an optionally substituted 5-8 membered ring including optionally substituted C3-C7 heterocycloalkyl including an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted C2-C7 heteroaryl including an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S; R'⁶, R'⁷, N'², and C'² together form an optionally substituted 5-8 membered ring including optionally substituted C3-C7 heterocycloalkyi including an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted C2-C7 heteroaryl including an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S; or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XII-1 ):

pharmaceutically acceptable salt thereof.

In some embodiments, the compound described by formula (XIII):

E' E

L'¹ L¹

I I

M2— A2— L A1 M1

(XIII)

wherein A1 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M1 ; A2 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M2; E¹ is a first monosaccharide or oligosaccharide moiety; E'¹ is a second monosaccharide or oligosaccharide moiety; L, L¹ , and L'¹ are remainders of L'; or a pharmaceutically acceptable salt thereof. In some embodiments, the compound is described by formula (XIII-1 ) :

(XIII-1 )

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g CH2CH(CH3)2), or cyclohexylmethyl; each of R⁶ and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each of R² and R'² is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2- C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XIII-2):

pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XIII-3):

(XIII-3)

pharmaceutically acceptable salt thereof.

In some embodiments, the compound described by formula (XIV):

E

L

M2— A2— L A1 M1

(XIV)

wherein A1 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M1 ; A2 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M2, or is absent; L and L¹ are remainders of L'; or a pharmaceutically acceptable salt thereof.

In some embodiments of the compounds described herein, R² is optionally substituted C1 -C5 alkamino. In some embodiments of the compounds described herein, R'² is optionally substituted C1 -C5 alkamino. In some embodiments, the optionally substituted C1 -C5 alkamino is CH2NH2 or CH2CH2NH2.

In some embodiments of the compounds described herein, R² is a polar moiety. In some embodiments of the compounds described herein, R'² is a polar moiety. In some embodiments of the compounds described herein, R⁶ is a polar moiety. In some embodiments of the compounds described herein, R'⁶ is a polar moiety. In some embodiments, the polar moiety includes a hydroxyl group, a carboxylic acid group, an ester group, or an amide group. In some embodiments, the polar moiety is hydroxyl substituted C1 -C4 alkyl. In some embodiments, the polar moiety is CH(CH3)OH or CH2OH.

In some embodiments of the compounds described herein, R⁸ is optionally substituted C1 -C5 alkamino. In some embodiments of the compounds described herein, R'⁸ is optionally substituted C1 -C5 alkamino. In some embodiments, the optionally substituted C1 -C5 alkamino is CH2NH2 or CH2CH2NH2. In some embodiments of the compounds described herein, R⁸ is optionally substituted C5-C1 5 aryl. In some embodiments of the compounds described herein, R'⁸ is optionally substituted C5-C15 aryl. In some embodiments, the optionally substituted C5-C15 aryl is naphthyl.

In some embodiments of the compounds described herein, R⁶, R⁷, N², and C² together form a 5- or 6-membered ring including C4-C5 heterocycloalkyl including an N heteroatom and additional 0 or 1 heteroatom independently selected from N, O, and S; and wherein R'⁶, R'⁷, N'², and C'² together form a 5- or 6-membered ring including C4-C5 heterocycloalkyl including an N heteroatom and additional 0 or 1 heteroatom independently selected from N, O, and S. In some embodiments, each of R² and R'² is C1 - C4 heteroalkylene. In some embodiments, each of R² and R'² is -(CH2)2NH- or -CH2NH-.

In some embodiments, the compound is described by formula (XV):

(XV)

pharmaceutically acceptable salt thereof.

In some embodiments of the compounds described herein, L', L, L¹ , or L'¹ includes one or more optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, optionally substituted C2-C15 heteroarylene, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino, wherein R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C1 5 heteroaryl.

In some embodiments of the compounds described herein, the backbone of L', L, L¹ , or L'¹ consists of one or more optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20

heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20

heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20

heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, optionally substituted C2-C15

heteroarylene, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino, wherein R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi , optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl , optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl , optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl , optionally substituted C8-C20 heterocycloalkynyl , optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl .

In some embodiments of the compounds described herein, L', L, L¹ , or L'¹ is oxo substituted. In some embodiments of the compounds described herein, the backbone of L', L, L¹ , or L'¹ includes no more than 250 atoms.

In some embodiments of the compounds described herein, L', L, L¹ , or L'¹ is capable of forming an amide, a carbamate, a sulfonyl, or a urea linkage.

In some embodiments of the compounds described herein, L, L¹ , or L'¹ is a bond.

In some embodiments of the compounds described herein, L is described by formula (L-l) :

L^c

L^B-Q-L^A

(L-l)

wherein L^A is described by formula G^A1-(Z^A1)_gi-(Y^A1)hi-(Z^A2)ii-(Y^A2)ji -(Z^A3)ki -(Y^A3)ii -(Z^A4)_mi -(Y^A4)ni-(Z^A5)oi - G^A2; L^B is described by formula G^B1-(Z^B1)g₂-(Y^B1

L^c is described by

G^A1 is a bond attached to Q in formula (L-l) ; G^A2 is a bond attached to A1 or M1 if A1 is absent; G^B1 is a bond attached to Q in formula (L-l) ; G^B2 is a bond attached to A2 or M2 if A2 is absent; G^C1 is a bond attached to Q in formula (L-l) ; G^C2 is a bond attached to E; each of Z^M , Z^A2, Z^A3, Z^M, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4, and Z^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 - C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20

heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; each of Y^A1 , Y^A2, Y^A3, Y^A4, Y^B1 , Y^B2, Y^B3, Y^B4, Y^C1 , Y^C2, Y^C3, and Y^C4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl , phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl , optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C1 5 heteroaryl; each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, and o3 is, independently, 0 or 1 ; Q is a nitrogen atom, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene.

In some embodiments of the compounds described herein, L is

wherein R^* is a bond or includes one or more of optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2- C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, optionally substituted C2-C15 heteroarylene, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, and imino, and wherein R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2- C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl.

In some embodiments of the compounds described herein, L is described by formula (L-lla) or formula (L-llb):

(L-lla) (L-llb)

wherein L^A is described by formula G^A1-(Z^A1)_gi-(Y^A1)hi-(Z^A2)ii-(Y^A2)ji -(Z^A3)ki -(Y^A3)ii -(Z^A4)_mi -

(Y^A4)m-(Z^A5)oi-G^A2; L^B is described by formula G^B1-(Z^B1 )_g2-(Y^B1 )h2-(Z^B2)i2-(Y^B2)j2-(Z^B3)_k2-(Y^B3)i2-(Z^B4)m2-

(Y^B4)n2-(Z^B5)_o2-G^B2; L^c is described by formula G^c1-(Z^c1 )_g3-(Y^c1 )h3-(Z^C2)i₃-(Y^C2)j3-(Z^C3)k3-(Y^C3)i3-(Z^C4)m3- (Y^C4)n3-(Z^C5)o3-G^C2; L^D is described by formula G^D1 -(Z^D1 )_g4-(Y^D1 )h4-(Z^D2)i₄-(Y^D2)j4-(Z^D3)k4-(Y^D3)i4-(Z^D4)m4-

(Y^D4)_n4-(Z^D5)o4-G^D2; G^A1 is a bond attached to N^A in formula (L-lla) or N^A in formula (L-llb); G^A2 is a bond attached to A1 or M1 if A1 is absent; G^B1 is a bond attached to N^A in formula (L-lla) or N^A in formula (L- llb); G^B2 is a bond attached to A2 or M2 if A2 is absent; G^C1 is a bond attached to N^B in formula (L-lla) or C in formula (L-llb); G^C2 is a bond attached to a first monosaccharide or oligosaccharide moiety, E¹ ; G^D1 is a bond attached to N^B in formula (L-lla) or C in formula (L-llb); G^D2 is a bond attached to a second monosaccharide or oligosaccharide moiety, E'¹ ; each of Z^A1 , Z^A2, Z^A3, Z^A4, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4, Z^C5, Z^D1 , Z^D2, Z^D3, Z^D4, and Z^D5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; each of Y^A1 , Y^A2, Y^A3, Y^A4, Y^B1 , Y^B2, Y^B3, Y^B4, Y^C1 , Y^C2, Y^C3, Y^C4, Y^D1 , Y^D2, Y^D3, and Y^D4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2- C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; and each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, o3, g4, h4, i4, j4, k4, I4, m4, n4, and o4 is, independently, 0 or 1 .

In some embodiments of the compounds described herein, L is

In some embodiments of the compounds described herein (e.g., compound of any one of formulas (XIII) and (XIV)), L is described by formula (L-lll):

H^A1 -(W^A1 )_gi -(X^A1 )hi -(W^A2)ii -(X^A2)ji -(W^A3)ki -(X^A3)ii -(W^A )_mi -(X^A4)m -(W^A5)₀1 -H^A2

(L-lll) wherein H^A1 is a bond attached to A2; H^A2 is a bond attached to A1 ; each of W^A1 , W^A2, W^A3, W^M , and W^A5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene; each of X^A1 , X^A2, X^A3, X^M is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C1 5 heteroaryl; and each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , and o1 is, independently, 0 or 1 . In some embodiments, L is

In some embodiments of the compounds described herein (e.g., compound of any one of formulas (XIII) and (XIV)), L¹ is described by formula (L-IV):

H^B1-(W^B1 )g2-(X^B1 )h2-(W^B2)i2-(X^B2)j2-(W^B3)k2-(X^B3)l2-(W^B4)m2-(X^B4)n2-(W^B5)o2-H^B2

(L-IV)

wherein H^B1 is a bond attached to A1 ; H^B2 is a bond attached to a first monosaccharide or oligosaccharide moiety, E¹ ; each of W^B1 , W^B2, W^B3, W^B4, and W^B5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4- C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; each of X^B1 , X^B2, X^B3, X^B4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 - C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20

cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; and each of g2, h2, i2, j2, k2, I2, m2, n2, and o2 is,

independently, 0 or 1 . In some embodiments, L¹ is

In some embodiments of the compounds described herein (e.g., compound of any one of formulas (XIII) and (XIV)), L'¹ is described by formula (L-V):

H^C1 -(W^C1 )g₃-(X^C1 )h3-(W^C2)_i3-(X^C2)j3-(W^C3)k3-(X^C3)l3-(W^C4)m3-(X^C4)n3-(W^C5)o3H^C2

(L-V)

wherein H^C1 is a bond attached to A2; H^C2 is a bond attached to a second monosaccharide or oligosaccharide moiety, E'¹ ; each of W^C1 , W^C2, W^C3, W^C4, and W^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; each of X^C1 , X^C2, X^C3, X^C4 is, independently, O, S, NR\ P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C1 5 heteroaryl ; and each of g3, h3, i3, j3, k3, I3, m3, n3, and o3 is, independently, 0 or 1 . In some embodiments, L'¹ is

In some embodiments of the compounds described herein, E, E¹ , or E'¹ is

In some embodiments, E, E¹ , or E'¹ is

In some embodiments, E, E¹ , or E'¹ is any one of the moieties in Tables 2A and 2B.

In some embodiments, E, E¹ , or E'¹ directly or indirectly activates an immune cell.

In some embodiments, E, E¹ , or E'¹ is a ligand to an innate immune receptor. In some embodiments, the innate immune receptor is AICL, BDCA2, CLEC2, Complement receptor 3,

Complement receptor 4, DCIR, dectin-1 , dectin-2, DC-SIGN, a C-Type lectin receptor, MMR, langerin, TLR2, Mincle, MBL, or KCR.

In some embodiments, E, E¹ , or E'¹ binds to an antibody. In some embodiments, the antibody is a natural antibody. In some embodiments, the natural antibody is an antibody of the immunoglobulin M (IgM) isotype. In some embodiments, the antibody binds to a moiety in Tables 2A and 2B. In some embodiments, the antibody is anti-aGal antibody or anti-aRha antibody.

In some embodiments, the monosaccharide moiety has one optionally substituted C6-C9 monosaccharide residue.

In some embodiments, the oligosaccharide moiety has 2-18 optionally substituted C6-C9 monosaccharide residues. In some embodiments, the oligosaccharide moiety has 2-12 optionally substituted C6-C9 monosaccharide residues. In some embodiments, each of the optionally substituted C6-C9 monosaccharide residues is, independently, glucose (Glc), galactose (Gal), mannose (Man), allose (All), altrose (alt), gulose (Gul), idose (ido), talose (Tal), fucose (Fuc), rhamnose (Rha or L-Rha), thia-rhamnose (thia-Rha or thia-L-Rha), quinovose (Qui), 2-deoxyglucose (2-dGlc), glucosamine (GlcN), galactosamine (GaIN), mannosamine (ManN), fucosamine (FucN), quinovosamine (QuiN), N-Acetyl- glucosamine (GlcNAc), N-Acetyl-galactosamine (GalNAc), N-Acetyl-mannosamine (ManNAc), N-acetyl- fucosamine (FucNAc), N-acetyl-quinovosamine (QuiNAc), glucuronic acid (GIcA), galacturonic acid (GalA), mannuronic acid (ManA), iduronic acid (IdoA), sialic acid (Sia), neuraminic acid (Neu), N-Acetyl- neuraminic acid (Neu5Ac), N-Glycolyl-neuraminic acid (Neu5Gc), glucitol (Glc-ol), galactitol (Gal-ol), mannitol (Man-ol), fructose (Fru), sorbose (Sor), tagatose (Tag), thevetose (The), acofriose (Aco), digitoxose (Dig), cymarose (Cym), abequose (Abe), colitose (Col), tyvelose (Tyv), ascarylose (Asc), paratose (Par), or N-acetyl-muramic acid (MurNAc).

In some embodiments, each of the optionally substituted C6-C9 monosaccharide residues is, independently, an optionally substituted C6 monosaccharide residue.

In some embodiments, the optionally substituted C6 monosaccharide residue is Glc, Gal, Man, All, Alt, Gul, Ido, or Tal. In some embodiments, the optionally substituted C6 monosaccharide residue is Fuc, Rha or L-Rha, thia-Rha or thia-L-Rha, Qui, or 2-dGlc. In some embodiments, the optionally substituted C6 monosaccharide residue is GlcN, GaIN, ManN, FucN, or QuiN. In some embodiments, the optionally substituted C6 monosaccharide residue is N- GlcNAc, GalNAc, ManNAc, FucNAc, QuiNAc, GIcA, GalA, ManA, or IdoA. In some embodiments, the optionally substituted C6 monosaccharide residue is Glc-ol, Gal-ol, or Man-ol. In some embodiments, the optionally substituted C6 monosaccharide residue is Fru, Sor, Tag. In some embodiments, the optionally substituted C6 monosaccharide residue is The, Aco, Dig, Cym, Abe, Col, Tyv, Asc, Par, or MurNAc. In some embodiments, the optionally substituted C6 monosaccharide residue is Rha, Gal, Glc, GlcA, GlcNAc, GalNAc, GlcN(Gc) (N-Glycolyl-Glucosamine), Neu5Ac, Neu5Gc, Fuc, Man, -hkPCbMan (mannose phosphate), 6-H2PO3GIC (glucose phosphate), Mur (muramoyl), Mur-L-Ala-D-i-Gln-Lys, (Mur)-3-0-GlcNAc, sulfate-galactose (Su-Gal), disulfate-galactose (Su₂-Gal), sulfate-glucose (Su-Glc), sulfate-GlcNAc (Su-GlcNAc), or sulfate-GalNAc (Su-GalNAc).

In some embodiments, the optionally substituted C6 monosaccharide residue is optionally substituted Rha. In some embodiments, the optionally substituted C6 monosaccharide residue is Rha.

In some embodiments, the optionally substituted C6 monosaccharide residue is L-Rha.

In some embodiments, the optionally substituted C6 monosaccharide residue is optionally substituted Gal or optionally substituted Glc. In some embodiments, the optionally substituted Gal is optionally substituted a1 -3Gal. In some embodiments, the optionally substituted Glc is an optionally substituted β-glucan having 1 -6 Glc moieties.

In some embodiments, the optionally substituted β-glucan is

pustulan.

In some embodiments, the optionally substituted β-glucan is laminarin.

In some embodiments, each of the optionally substituted C6-C9 monosaccharide residues is, independently, an optionally substituted C9 monosaccharide residue, wherein the optionally substituted C9 monosaccharide residue is Sia, Neu, Neu5Ac, or Neu5Gc.

In some embodiments, at least one optionally substituted C6-C9 monosaccharide residue is substituted with 1 -3 substituents independently selected from sulfate, phosphate, methyl, acetyl, natural amino acids, and non-natural amino acids. In some embodiments, at least one optionally substituted C6- C9 monosaccharide residue is substituted with 1 -3 substituents independently selected from sulfate, phosphate, methyl, and acetyl. In some embodiments, at least one optionally substituted C6-C9 monosaccharide moiety is substituted with 1 -3 substituents independently selected from natural and non- natural amino acids. In some embodiments, the natural amino acid is alanine, lysine, serine, glutamine or asparagine.

In another aspect, the disclosure features a compound of formula (XVI):

(XVI)

wherein each A is an independently selected amino acid; L is a linker that, when m is 2, 3, 4, or 5, is bound to any of A; each E is independently selected from a monosaccharide or an oligosaccharide; m is 0, 1 , 2, 3, 4, or 5; n is 1 , 2, 3, 4, or 5; and Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVI), Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of a natural amino acid, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVI), at least one of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof. In some embodiments, at least two of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof. In some embodiments, at least three of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof. In some embodiments, at least four of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof. In some embodiments, at least five of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVI), each Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of serine, threonine, cysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVI), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVI), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XVII):

(XVII)

wherein each A¹ and A² is an independently selected amino acid; L is a linker that, when m is 1 , 2, 3, 4, or 5, is bound to a nitrogen atom in any A¹ and a nitrogen atom in any A²; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; n is 1 , 2, 3, 4, or 5; and Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each

independently selected from the side chain of a natural amino acid, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), at least one (e.g., at least two, three, four, or five) of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), each Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of serine, threonine, cysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyl, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XVII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

In some embodiments of a compound of formula (XVII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; and each m is independently selected from 1 , 2, 3, 4, or 5; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), each m is independently 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

In some embodiments of a compound of formula (XVII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; each m is independently 1 , 2, 3 or 4; E is a monosaccharide; n is 1 , 2, 3, or 4; and the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

In another aspect, the disclosure features a compound of formula (XVIII):

(XVIII)

wherein each A¹ and A² is an independently selected amino acid; X is absent or is -CH2CH2C(0)NH-; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; n is 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 10; and e is an integer from 0 to 10; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVIII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVIII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVIII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2 or 3; d is an integer from 1 to 10; e is an integer from 1 to 10; and E is a monosaccharide; or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XVIII), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; m is 2 or 3; d is an integer from 1 to 10; e is an integer from 1 to 10; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments, m is 2. In some embodiments, m is 3.

In some embodiments of a compound of formula (XVIII), d is 1 0; e is 10; and X is absent; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVIII), m is 2; d is 1 ; e is 1 ; and X is - CH2CH2C(0)NH-; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XVIII), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

, or a

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XIX):

(XIX)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; n is 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 10; e is an integer from 0 to 10; f is an integer from 0 to 10; and g is an integer from 0 to 10; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XIX), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XIX), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and

Q⁶ is selected from one of

or a p armaceutca y accepta e sat t ereo . In some embodiments of a compound of formula (XIX), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

In some embodiments of a compound of formula (XIX), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2; d is 1 ; e is 1 ; f is 3; g is 1 ; and E is a monosaccharide; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XIX), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; m is 2; d is 1 ; e is 1 ; f is 3; g is 1 ; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XIX), E is

pharmaceutically acceptable salt thereof. In some embodiments, E i

s or a

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XX):

(XX) wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 1 0; e is an integer from 0 to 10; f is an integer from 0 to 10; and g is an integer from 0 to 10; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XX), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XX), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

In some embodiments of a compound of formula (XX), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2; d is 1 ; e is 1 ; f is 1 ; g is 1 ; and E is a monosaccharide; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XX), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; m is 2; d is 1 ; e is 1 ; f is 1 ; g is 1 ; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XX), E is

pharmaceutically acceptable salt thereof. In some embodiments, E

is or a

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula

(XXI)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 1 0; e is an integer from 0 to 10; f is an integer from 0 to 10; g is an integer from 0 to 25; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXI), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXI), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XXI), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2; d is 1 ; e is 1 ; f is 1 ; g is 8 to 25; and E is a monosaccharide; or a

pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXI), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; m is 2; d is 1 ; e is 1 ; f is 1 ; g is 8 to 25; E is a

monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXI), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXII):

(XXII)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; and d is an integer from 0 to 15; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2; d is 10; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXII), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and piperazine-2-carboxylic acid; m is 2; d is 10; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXII), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

pharmaceutically acceptable salt thereof. In another aspect, the disclosure features a compound of formula (XXIII):

(XXIII)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from

a monosaccharide or an oligosaccharide; Y is

, -C(0)CH2CH2-, -CH2-, or is absent; X is -C(0)CH2CH2- or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 15; e is an integer from 1 to 10; f is an integer from 1 to 5; and g is an integer from 1 to 5; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- -5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alph

neopentylglycine; m is 2, 3 or 4; d is 1 ; when Y is

, e is 4; E is a monosaccharide; f is 1 or 2; g is 1 or 2; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 3-aminoalanine, 2-piperazinecarboxylic acid, 2-aminohexanoic

acid, 2-aminooctanoic acid, methionine, and threonine; m is 2, 3 or 4; when Y is

, e is 4; d is 1 ; E is a monosaccharide; f is 1 or 2; g is 1 or 2; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 3 or 4; d is 1 ; Y is absent; E is a monosaccharide; f is 1 ; g is 1 ; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; and m is 3; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3 or 4; d is 1 ; Y is -C(0)CH2CH2-; E is a monosaccharide; f is 1 or 2; g is 1 or 2; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments, each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 2,4- diaminobutyric acid, 3-aminoalanine, 2-aminohexanoic acid, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S- propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5-methylhexanoic acid, 2-amino-5- methylhex-4-enoic acid, alpha-t-butylglycine, and alpha-neopentylglycine, or a pharmaceutically acceptable salt thereof. In some embodiments, m is 4, d is 1 , f is 1 , and g is 1 , or a pharmaceutically acceptable salt thereof. In some embodiments, m is 3, f is 2, and g is 1 , or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3 or 4; d is 1 ; Y is -CH2-; E is a monosaccharide; f is 1 or 2; g is 1 or 2; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments, each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2- aminooctanoic acid and threonine; f is 1 ; and g is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments, each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2- aminohexanoic acid, 2-aminooctanoic acid and threonine, or a pharmaceutically acceptable salt thereof; m is 4; f is 1 ; and g is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments, each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminohexanoic acid, 2-aminooctanoic acid and threonine, or a pharmaceutically acceptable salt thereof; m is 3; f is 1 ; and g is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIII), E is ; or a

pharmaceutically acceptable salt thereof. In some embodiments, E is

, or a

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXIV):

(XXIV)

wherein each A¹ and A² is an independently selected amino acid; X is -C(0)CH2CH2CH2-Y- or - C(0)CH2CH2C(0)NH-; Y is heteroaryl; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; and d is an integer from 0 to 15; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIV), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIV), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIV), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2 or 3; d is an integer from 1 to 3; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIV), each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 3-(2-naphthyl)alanine, and threonine, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIV), X is -C(0)CH2CH2CH2-Y-; m is 3; d is 3; and Y is 1 ,4-triazololyl; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIV), X is -C(0)CH2CH2C(0)-; m is 2; and d is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIV), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXV):

(XXV)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 1 5; and e is an integer from 0 to 1 5; or a

pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXV), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XXV), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

In some embodiments of a compound of formula (XXV), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

In some embodiments of a compound of formula (XXV), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2 or 3; d is an integer from 1 to 3; e is an integer from 1 to 3; E is a

monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXV), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; m is 2 or 3; d is an integer from 1 to 3; e is an integer from 1 to 3; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXV), E is or a

pharmaceutically acceptable salt thereof. In some embodiments, E i

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXVI):

(XXVI) wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; R is C1 -C20 alkyl; each m is independently 0, 1 , 2, 3, 4, or 5;each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 20; e is an integer from 0 to 20; and f is an integer from 0 to 20; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVI), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVI), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVI), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2 or 3; d is an integer from 1 to 3; e is an integer from 1 to 3; f is an integer from 1 to 3; E is a monosaccharide; R is C1 -C1 0 alkyl ; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVI), each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; m is 2; d is 1 ; e is 1 ; f is 1 ; E is a monosaccharide; R is C1 -C1 0 alkyl ; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XXVI), E is or a

pharmaceutically acceptable salt thereof. In some embodiments, E is

or a

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXVII)

(XXVII)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; R is C1 -C20 alkyl; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 20; e is an integer from 1 to 20; and f is an integer from 1 to 20; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceutca y accepta e sat t ereo .

In some embodiments of a compound of formula (XXVII), the combination of Q¹, Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

In some embodiments of a compound of formula (XXVII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3, or 4; d is an integer from 1 to 3; e is an integer from 1 to 3; f is an integer from 1 to 3; E is a monosaccharide; R is C1 -C10 alkyl; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVII), each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; m is 2, 3, or 4; d is 1 ; e is 1 ; f is 1 ; E is a monosaccharide; R is C1 -C10 alkyl; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments, m is 2. In some embodiments, m is 3. In some embodiments, m is 4.

In some embodiments of a compound of formula (XXVII), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXVIII):

a monosaccharide or an oligosaccharide; Y is

-C(0)CH2CH2-, -CH2-, or is absent; e is an integer from 1 to 10; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; and d is an integer from 0 to 15; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVIII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVIII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

or a p armaceut ca y accepta e sa t t ereo .

In some embodiments of a compound of formula (XXVIII), Y is -C(0)CH2CH2-, or a

pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVIII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3, or 4; d is an integer from 1 to 3; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVIII), each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; and d is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXVIII), m is 2. In some embodiments of a compound of formula (XXVIII), m is 3. In some embodiments of a compound of formula (XXVIII), m is 4. In some embodiments of a compound of formula (XXVIII), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

or a

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXIX):

(XXIX)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; X is -CH2- or -C(O)-; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; and d is an integer from 0 to 15; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIX), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIX), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIX), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3, or 4; d is an integer from 1 to 3; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XXIX), each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; and d is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXIX), m is 2. In some embodiments of a compound of formula (XXIX), m is 3. In some embodiments of a compound of formula (XXIX), m is 4.

In some embodiments of a compound of formula (XXIX), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXX):

(XXX)

a monosaccharide or an oligosaccharide; Y is

-C(0)CH2CH2-, -CH2-, or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 15; g is an integer from 0 to 15; and e and e' are each independently an integer from 1 to 3; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXX), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXX), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXX), Y is -C(0)CH2CH2-, or a

pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXX), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2- carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2- naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4- amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S- methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5- methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3, or 4; d is an integer from 1 to 3; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXX), each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; and d is 1 ; and e is 1 or 2; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXX), m is 2. In some embodiments of a compound of formula (XXX), m is 3. In some embodiments of a compound of formula (XXX), m is 4. pharmaceutically acceptable salt thereof. In some embodiments, E is

, or a pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXXI) :

(XXXI)

a monosaccharide or an oligosaccharide; each Y is independently selected from

C(0)CH2CH2-, and -CH2-, or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 1 5; each e is independently from 1 to 1 5; each f is independently from 1 to 1 5; and g is from 1 to 1 5; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXI), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl , 2-butyl , methyl , and 2-aminoethyl , or a pharmaceutically acceptable salt thereof. In some embodiments of a compound of formula (XXXI), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

In some embodiments of a compound of formula (XXXI), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

In some embodiments of a compound of formula (XXXI), Y is -C(0)CH2CH2-, or a

pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXI), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3, or 4; d is an integer from 1 to 5; e is an integer from 1 to 5; E is a

monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments, each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; and d is 4 or 5; e is 4 or 5; and f is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXI), m is 2. In some embodiments of a compound of formula (XXXI), m is 3. In some embodime m is 4.

In some embodiments of a compound of formula (XXXI),

pharmaceutically acceptable salt thereof. In some embod

iments, E is , or a

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXXII):

(XXXII)

a monosaccharide or an oligosaccharide; each Y is independently selected from

C(0)CH₂CH2-, -CH2CH2NHC(0)CH₂CH2-, and -CH₂-, or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 15; each e is independently from 1 to 15; and g is from 1 to 15; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXII), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXII), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXII), Y is -CH2CH2NHC(0)CH2CH2-, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXII), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4- phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2- aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2- amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha- neopentylglycine; m is 2, 3, or 4; d is an integer from 1 to 5; e is an integer from 1 to 5; E is a

monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXII), each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; and d is an integer from 1 to 4; and e is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXII), d is 2. In some embodiments of a compound of formula (XXXII), m is 2. In some embodiments, m is 3. In some embodiments, m is 4.

In some embodiments of a compound of formula (XXXII), E is

pharmaceutically acceptable salt thereof. In some embodiments, E is

pharmaceutically acceptable salt thereof.

In another aspect, the disclosure features a compound of formula (XXXIII):

(XXXIII)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; each X is independently selected from

, -C(0)CH₂CH₂C(0)-, -CH2CH2NHC(0)CH₂CH2-, -C(O)-, and -CH₂-, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 alkenyl, optionally substituted C1 -C20 alkynyl, optionally substituted C1 -C15 aryl, optionally substituted C1 -C15 heteroaryl, or is absent; each Y is independently selected from -CH-, or oxygen; Z is -NH-, -NHC(O)-, oxygen, or sulfur; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 15; g is an integer from 1 to 15; e and e' are independently from 1 to 10 and or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXIII), d is 2.

In some embodiments of a compound of formula (XXXIII), the compound is of the formula (XXXIII-1 ):

(XXXIII-1 )

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXIII), the compound is of the formula (XXXI 11-2):

(XXXIII-2)

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXIII) (e.g., a compound of formula (XXXIII-1 ) or a compound of formula (XXXIII-2)), each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof. In some embodiments, each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2- methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl. hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXIII) (e.g., a compound of formula (XXXIII-1 ) or a compound of formula (XXXIII-2)), the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceutca y accepta e sat t ereo .

In some embodiments of a compound of formula (XXXIII) (e.g., a compound of formula (XXXII 1- ) or a compound of formula (XXXIII-2)), the combination of Q¹, Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceut ca y accepta e sa t t ereo .

In some embodiments of a compound of formula (XXXIII) (e.g., a compound of formula (XXXII 1- ) or a compound of formula (XXXIII-2)), each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4- diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2-naphthyl)alanine, 2- piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5-methylhexanoic acid, 2-amino-5- methylhex-4-enoic acid, alpha-t-butylglycine, and alpha-neopentylglycine; m is 2, 3, or 4; d is an integer from 1 to 5; e is 1 ; X is -C(0)CH2CH2C(0)-; E is a monosaccharide; and n is 1 ; or a pharmaceutically acceptable salt thereof. In some embodiments, each A¹ and A² is independently selected from 2,4- diaminobutyric acid, 2-aminohexanoic acid, and threonine; and d is 1 ; or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXIII) (e.g., a compound of formula (XXXII 1- ) or a compound of formula (XXXIII-2)), m is 2. In some embodiments, m is 3. In some embodiments, m is 4. In some embodiments of a compo (e.g., a compound of formula (XXXII 1-1 ) or a

compound of formula (XXXIII-2)), or a pharmaceutically acceptable salt

In some embodiments, E

ically acceptable salt thereof.

In some embodiments, compound is described by formula (XXXIV)

(XXXIV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH₂CH(CH₃)2), or cyclohexylmethyl; each of R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R'², R'³, R'⁴, R'⁵, R'⁶, R'⁷, R'⁸, R'⁹, and R'¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; or two R groups on the same or adjacent atoms join to form an optionally substituted 5-8 membered ring;

each of a', b', c', a, b, and c is, independently, 0 or 1 ;

each of N¹ , N², N³, N⁴, N'¹ , N'², N'³, and N'⁴ is a nitrogen atom;

each of C¹ , C², C³, C'¹ , C'², and C'³ is a carbon atom ;

L is a linker comprising at least one optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C3-C15 heteroarylene;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound comprises at least one optionally substituted 5-8 membered ring formed by joining (i) R², R³, and C¹ ; (ii) R³, R⁴, N¹ , and C¹ ; (iii) R⁵, R⁶, and C²; (iv) R⁶, R⁷, N², and C²; (v) R⁸, R⁹, and C³; (vi) R⁹, R¹⁰, N³, and C³; (vii) R'², R'³, and C'¹ ; (viii) R'³, R'⁴, N'¹ , and C'¹ ; (ix) R'⁵, R'⁶, and C'²; (x) R'⁶, R'⁷, N'², and C'²; (xi) R'⁸, R'⁹, and C'³; or (xii) R'⁹, R'¹⁰, N'³, and C'³.

In some embodiments, L is described by formula (L-VI) :

L^c

L^B-Q-L^A

(L-VI)

wherein L^A is described by formula G^A1-(Z^A1)_gi-(Y^A1)hi-(Z^A2)ii-(Y^A2)ji-(Z^A3)ki-(Y^A3)ii-(Z^A4)_mi-

L^B is described by formula G^B1-(Z^B1 )_g2-(Y^B1 )h2-(Z^B2)i2-(Y^B2)j2-(Z^B3)_k2-(Y^B3)i2-(Z^B4)m2-(Y^B4)n2-(Z^B5)02-

G^B2;

L^c is described by formula G^c1-(Z^{c 1} )_g3-(Y^c1 )h3-(Z^C2)i₃-(Y^C2)j3-(Z^C3)k3-(Y^C3)i3-(Z^C4)m3-(Y^C4)n3-(Z^C5)03-

G^C2;

G^A1 is a bond attached to Q in formula (L-VI);

G^A2 is a bond attached to A1 or M1 if A1 is absent;

G^B1 is a bond attached to Q in formula (L-VI);

G^B2 is a bond attached to A2 or M2 if A2 is absent;

G^C1 is a bond attached to Q in formula (L-VI);

G^C2 is a bond attached to E; each of Z^A1 , Z^A2, Z^A3, Z^A4, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4 and Z^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene;

each of Y^A1 , Y^A2, Y^A3, Y^A4, Y^B1 , Y^B2, Y^B3, Y^B4, Y^C1 , Y^C2, Y^C3, and Y^C4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino;

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, and o3 is, independently, 0 or 1 ;

Q comprises at least one optionally substituted C3-C20 cycloalkylene, optionally substituted C3- C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C3-C15 heteroarylene.

In some embodiments, L is described b formula (L-VII):

-Q- wherein L^A is described by formula G^A1-(Z^A1)_gi-(Y^A1)hi-(Z^A2)ii-(Y^A2)ji -(Z^A3)ki -(Y^A3)ii -(Z^A4)_mi -

G^B2;

L^c is described by formula G^c1 -(Z^c1 )_g3-(Y^c1 )h3-(Z^C2)i₃-(Y^C2)j3-(Z^C3)k3-(Y^C3)i3-(Z^C4)m3-(Y^C4)n3-(Z^C5)03-

G^C2;

L^D is described by formula G^D1 -(Z^D1 )_g4-(Y^D1 )h4-(Z^D2)i₄-(Y^D2)j4-(Z^D3)k4-(Y^D3)i4-(Z^D4)m4-(Y^D4)n4-(Z^D5)o4-

G^D2;

G^A1 is a bond attached to Q in formula (L-VII);

G^A2 is a bond attached to A1 or M1 if A1 is absent; G^B1 is a bond attached to Q in formula (L-VII);

G^B2 is a bond attached to A2 or M2 if A2 is absent;

G^C1 is a bond attached to Q in formula (L-VII);

G^C2 is a bond attached to a first monosaccharide or oligosaccharide moiety, E¹ ;

G^D1 is a bond attached to N in formula (L-VII);

G^D2 is a bond attached to a second monosaccharide or oligosaccharide moiety, E²;

each of Z^A1 Z^A2 Z^A3 Z^A4 Z^A5 Z^B1 Z^B2 Z^B3 Z^B4 Z^B5 Z^C1 Z^C2 Z^C3 Z^C4 Z^C5 Z^D1 Z^D2 Z^D3 Z^D4 and Z^D5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20

heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20

heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene;

each of Y^A1 Y^A2 Y^A3 Y^A4 Y^B1 Y^B2 Y^B3 Y^B4 Y^C1 Y^C2 Y^C3 Y^C4 Y^D1 Y^D2 Y^D3 and Y^D4 is independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino;

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, o3, g4, h4, i4, j4, k4, I4, m4, n4, and o4 is, independently, 0 or 1 ;

In some embodiments the compound is described by formula (XXXV):

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH₂CH(CH₃)2), or cyclohexylmethyl;

each of R², R⁶, R⁸, R'², R'⁶, and R'⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments the compound is described by formula (XXXV):

(XXXV)

each of R², R⁶, R⁸, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXVI):

(XXXVI)

each of R², R⁶, R⁸, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments the compound is described by formula (XXXVII):

(XXXVII)

each of R², R⁶, and R⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXVIII):

(XXXVIII)

each of R², R⁶, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXIX):

(XXXIX)

each of R², R⁶, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments the compound is described by formula (XXXX):

(XXXX)

each of R² and R⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXXI):

(XXXXI) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH₂CH(CH₃)2), or cyclohexylmethyl;

each of R² and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments the compound is described by formula (XXXXII):

(XXXXII)

R² is a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXXIII):

(XXXXIII)

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

In some embodiments, L comprises at least one an optionally substituted C3 cycloalkylene or an optionally substituted C3 heterocycloalkylene.

In some embodiments the compound is described by formula (XXXXIV):

(XXXXIV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH₂CH(CH₃)2), or cyclohexylmethyl; and

L comprises an optionally substituted C3 cycloalkylene or an optionally substituted C3 heterocycloalkylene,

or a pharmaceutically acceptable salt thereof.

In some embodiments the compound is described by formula (XXXXV):

(XXXXV)

or a pharmaceutically acceptable salt thereof.

In some embodiments, L is

wherein each q is, independently, an integer from 1 to 1 1 , inclusive.

In some embodiments, L comprises at least one an optionally substituted C5 cycloalkyl optionally substituted C5 heterocycloalkylene.

In some embodiments, the compound is described by formula (XXXXVI):

(XXXXVI)

L comprises an optionally substituted C5 cycloalkylene or an optionally substituted C5 heterocycloalkylene, or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXXVII):

(XXXXVII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted

C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH₂CH(CH₃)2), or cyclohexylmethyl; and

L comprises an optionally substituted C5 cycloalkylene or an optionally substituted C5 heterocycloalkylene,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXXVIII):

(XXXXVIII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH₂CH(CH₃)2), or cyclohexylmethyl; and L comprises an optionally substituted C5 cycloalkylene or an optionally substituted C5 heterocycloalkylene,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is described by formula (XXXXIX):

(XXXXIX)

or a pharmaceutically acceptable salt thereof.

In some embodiments L is

wherein each q is, independently, an integer from 1 to 1 1 , inclusive.

In some embodiments, L comprises at least one an optionally substituted C6 cycloalkylene, at least one an optionally substituted C6 heterocycloalkylene, or at least one an optionally substituted C6 arylene.

In some embodiments the compound is described by formula (XXXXX):

(XXXXX)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH₂CH(CH₃)2), or cyclohexylmethyl; and L comprises an optionally substituted C6 arylene,

or a pharmaceutically acceptable salt thereof.

In some embodiments, L is

wherein each q is, independently, an integer from 1 to 1 1 , inclusive.

In some embodiments, non-natural amino acids that may be included in a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)), for example, in the polymyxin core portions (e.g., in the first cyclic heptapeptide and/or the second cyclic heptapeptide) and/or the linker portion of the compound, include, but are not limited to, D-Ser, D-Pro, D-Leu, D-Nle (D-norleucine), D-Thr, D-Val, L-Abu (L-2-aminobutyric acid), 3-(2H-tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4- diaminobutyric acid, 3-hydroxyproline, 2-amino-4-phenylbutyric acid, 3-(2-naphthyl)alanine, 2- piperazinecarboxylic acid, 2-aminooctanoic acid, 2-aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha-neopentylglycine. In some embodiments, polymyxin core portions (e.g., in the first cyclic heptapeptide and/or the second cyclic heptapeptide) of the compound include D-Ser, D- Pro, D-Leu, D-Nle, D-Thr, D-Val, and/or L-Abu.

In some embodiments of the compounds described herein, a concentration of the compound, or a pharmaceutically acceptable salt thereof, that activates an immune cell is less than or equal to 10,000 nM. In some embodiments, the concentration of the compound, or a pharmaceutically acceptable salt thereof, that activates an immune cell is less than or equal to equal to 1 ,000 nM. In some embodiments, the concentration of the compound, or a pharmaceutically acceptable salt thereof, that activates an immune cell is less than or equal to equal to 100 nM.

In another aspect, the disclosure features a pharmaceutical composition including a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition further includes an antibacterial agent.

In some embodiments, the antibacterial agent is selected from the group consisting of linezolid, tedizolid, posizolid, radezolid, retapamulin, valnemulin, tiamulin, azamulin, lefamulin, plazomicin, amikacin, gentamicin, gamithromycin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, streptomycin, spectinomycin, geldanamycin, herbimycin, rifaximin, loracarbef, ertapenem, doripenem, imipenem/cilastatin, meropenem, cefadroxil, cefazolin, cefalotin, cefalexin, cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepime, ceftaroline fosamil, ceftobiprole, teicoplanin, vancomycin, telavancin, dalbavancin, oritavancin, clindamycin, lincomycin, daptomycin, solithromycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin, aztreonam, furazolidone, nitrofurantoin, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin g, penicillin v, piperacillin, penicillin g, temocillin, ticarcillin, amoxicillin clavulanate, ampicillin/sulbactam,

piperacillin/tazobactam, ticarcillin/clavulanate, bacitracin, ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, nalidixic acid, norfloxacin, ofloxacin, trovafloxacin, grepafloxacin, sparfloxacin, temafloxacin, mafenide, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfadimethoxine, sulfamethizole, sulfamethoxazole, sulfanamide, sulfasalazine, sulfisoxazole, trimethoprim-sulfamethoxazole (tmp-smx), sulfonamidochrysoidine, eravacycline, demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline, clofazimine, dapsone, capreomycin, cycloserine, ethambutol(bs), ethionamide, isoniazid, pyrazinamide, rifampicin, rifabutin, rifapentine, streptomycin, arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin, quinupristin/dalfopristin, thiamphenicol, tigecycline, tinidazole, and trimethoprim, prodrugs thereof, and pharmaceutically acceptable salts thereof.

In some embodiments, a prodrug of tedizolid is tedizolid phosphate. In some embodiments, the antibacterial agent is tedizolid, azithromycin, meropenem, amikacin, levofloxacin, rifampicin, linezolid, erythromycin, or solithromycin. In some embodiments, the antibacterial agent is tedizolid, azithromycin, meropenem, amikacin, or levofloxacin.

In another aspect, the disclosure features a method of protecting against or treating a bacterial infection in a subject by administering to the subject a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)).

In some embodiments of the method, the method further includes administering to the subject an antibacterial agent.

In another aspect, the disclosure features a method of protecting against or treating a bacterial infection in a subject by administering to the subject (1 ) a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) and (2) an antibacterial agent.

In some embodiments of this aspect of the disclosure, the bacterial infection is caused by Gram- negative bacteria. In some embodiments, the bacterial infection is caused by a resistant strain of bacteria. In some embodiments, the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene and/or a chromosomal mutation conferring polymyxin resistance. In some embodiments, the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, or the mcr-3 gene. In some embodiments, the resistant strain of bacteria possesses a chromosomal mutation conferring polymyxin resistance. In some embodiments, the resistant strain of bacteria is a resistant strain of E. coli.

In another aspect, the disclosure features a method of protecting against or treating sepsis in a subject by administering to the subject a compound described herein (e.g., a compound of any one of formulas (I)- (XXXXX)).

In another aspect, the disclosure features a method of preventing lipopolysaccharides (LPS) in Gram- negative bacteria from activating an immune system in a subject by administering to the subject a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)). In some embodiments, the method prevents LPS from activating a macrophage. In some embodiments, the method prevents LPS-induced nitric oxide (NO) production from a macrophage. In some embodiments of this aspect of the disclosure, the Gram-negative bacteria is a resistant strain of Gram-negative bacteria. In some embodiments, the resistant strain of Gram-negative bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene and/or a chromosomal mutation conferring polymyxin resistance. In some embodiments, the resistant strain of Gram-negative bacteria possesses the mcr- 1 gene, the mcr-2 gene, or the mcr-3 gene. In some embodiments, the resistant strain of Gram- negative bacteria possesses a chromosomal mutation conferring polymyxin resistance. In some embodiments, the resistant strain of Gram-negative bacteria is a resistant strain of E. coli.

In some embodiments of this aspect of the disclosure, the method further includes administering to the subject an antibacterial agent.

In some embodiments, the compound and the antibacterial agent are administered substantially simultaneously.

In some embodiments, the compound and the antibacterial agent are administered separately. In some embodiments, the compound is administered first, followed by administering of the antibacterial agent alone. In some embodiments, the antibacterial agent is administered first, followed by administering of the compound alone.

In some embodiments, the compound and the antibacterial agent are administered substantially simultaneously, followed by administering of the compound or the antibacterial agent alone. In some embodiments, the compound or the antibacterial agent is administered first, followed by administering of the compound and the antibacterial agent substantially simultaneously.

In some embodiments, administering the compound and the antibacterial agent together lowers the MIC of each of the compound and the antibacterial agent relative to the MIC of each of the compound and the antibacterial agent when each is used alone.

In some embodiments of the methods described herein, the compound and/or the antibacterial agent is administered intramuscularly, intravenously, intradermal^, intraarterially, intraperitoneally, intralesionally, intracranial^, intraarticularly, intraprostatically, intrapleural^, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctival, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularally, orally, locally, by inhalation, by injection, or by infusion.

In another aspect, the disclosure features a method of preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria, by contacting the bacteria or a site susceptible to bacterial growth with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)).

In some embodiments, the method further includes contacting the bacteria or the site susceptible to bacterial growth with an antibacterial agent.

In another aspect, the disclosure features a method of preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria, including contacting the bacteria or a site susceptible to bacterial growth with (1 ) a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) and (2) an antibacterial agent.

In some embodiments of the methods described herein, the antibacterial agent is selected from the group consisting of linezolid, tedizolid, posizolid, radezolid, retapamulin, valnemulin, tiamulin, azamulin, lefamulin, plazomicin, amikacin, gentamicin, gamithromycin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, streptomycin, spectinomycin, geldanamycin, herbimycin, rifaximin, loracarbef, ertapenem, doripenem, imipenem/cilastatin, meropenem, cefadroxil, cefazolin, cefalotin, cefalexin, cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepime, ceftaroline fosamil, ceftobiprole, teicoplanin, vancomycin, telavancin, dalbavancin, oritavancin, clindamycin, lincomycin, daptomycin, solithromycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin, aztreonam, furazolidone, nitrofurantoin, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin g, penicillin v, piperacillin, penicillin g, temocillin, ticarcillin, amoxicillin clavulanate,

ampicillin/sulbactam, piperacillin/tazobactam, ticarcillin/clavulanate, bacitracin, ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, nalidixic acid, norfloxacin, ofloxacin, trovafloxacin, grepafloxacin, sparfloxacin, temafloxacin, mafenide, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfadimethoxine, sulfamethizole, sulfamethoxazole, sulfanamide, sulfasalazine, sulfisoxazole, trimethoprim-sulfamethoxazole (tmp-smx), sulfonamidochrysoidine, eravacycline, demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline, clofazimine, dapsone, capreomycin, cycloserine, ethambutol(bs), ethionamide, isoniazid, pyrazinamide, rifampicin, rifabutin, rifapentine, streptomycin, arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin, quinupristin/dalfopristin, thiamphenicol, tigecycline, tinidazole, and

trimethoprim, prodrugs thereof, and pharmaceutically acceptable salts thereof.

In some embodiments, the bacteria are Gram-negative bacteria. In some embodiments, the bacteria are a resistant strain of bacteria. In some embodiments, the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene and/or a chromosomal mutation conferring polymyxin resistance. In some embodiments, the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene. In some embodiments, the resistant strain of bacteria possesses a chromosomal mutation conferring polymyxin resistance. In some embodiments, the resistant strain of bacteria is a resistant strain of E. coli.

In another aspect, the disclosure features any compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX) and/or any one of compounds 1 -123, or a pharmaceutically

acceptablesalt thereof).

In some embodiments, the disclosure provides a compound described by any one Formulas (IV)- (XIII), (XV), or (XXXIV), wherein each of R'¹ and R¹ is, independently, benzyl or CH₂CH(CH₃)2.

Definitions

The term "cyclic heptapeptide" or "cycloheptapeptide," as used herein, refers to certain compounds that bind to lipopolysaccharides (LPS) in the cell membrane of Gram-negative bacteria to disrupt and permeabilize the cell membrane, leading to cell death and/or sensitization to other antibiotics. Cyclic heptapeptides or cycloheptapeptides comprise seven natural or non-natural a-amino acid residues, such as D- or L-amino acid residues, in a closed ring. Generally, cyclic heptapeptides are formed by linking the a-carboxyl group of one amino acid to the a-amino group or the γ-amino group of another amino acid and cyclizing. The cyclic heptapeptide comprises a heterocycle comprising carbon and nitrogen ring members, which may be substituted, for example, with amino acid side chains. One nitrogen from an a-amino group in the cyclic heptapeptide, however, is not a ring member and is branched from a ring member of the heterocycle. Thus, this nitrogen is directly attached to a ring member, such as a carbon atom (e.g., an a-carbon atom). This nitrogen atom serves as an attachment point for the cyclic heptapeptide to a linker and/or to a peptide (e.g., a peptide including 1 -5 amino acid residue(s)), and thus is referred to herein as a "linking nitrogen." The linking nitrogen is directly attached to the ring of the cyclic heptapeptide and is not derived from a side chain, such as an ethylamine side chain. The linking nitrogens in a compound of, e.g., formula (II) or (III), are N⁴ and N'⁴.

In some embodiments, a peptide including one or more (e.g., 1 -5; 1 , 2, 3, 4, or 5) amino acid residues (e.g., natural and/or non-natural amino acid residues) may be covalently attached to a linking nitrogen (e.g., N⁴ and/or N'⁴, the nitrogen from an a-amino group) in the cyclic heptapeptide ring. Cyclic heptapeptides may be derived from polymyxins (e.g., naturally existing polymyxins and non-natural polymyxins) and/or octapeptins (e.g., naturally existing octapeptins and non-natural octapeptins).

Examples of naturally existing polymyxins include, but are not limited to, polymyxin Bi , polymyxin B2, polymyxin B3, polymyxin B4, polymyxin B5, polymyxin Ββ, polymyxin BHIe, polymyxin B2-lle, polymyxin Ci , polymyxin C2, polymyxin Si , polymyxin Ti , polymyxin T2, polymyxin Ai , polymyxin A2, polymyxin Di , polymyxin D2, polymyxin Ei (colistin A), polymyxin E2 (colistin B), polymyxin E3, polymyxin E₄, polymyxin E7, polymyxin EHIe, polymyxin Ei-Val, polymyxin Ei-Nva, polymyxin E2-lle, polymyxin E2-Val, polymyxin E2-Nva, polymyxin Es-lle, polymyxin Mi , and polymyxin M2. In other embodiments, a cyclic heptapeptide may be entirely synthetic and prepared by standard peptide methodology as known in the art.

The term "covalently attached" refers to two parts of a compound that are linked to each other by a covalent bond formed between two atoms in the two parts of the compound. For example, in the compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)), when a' is 0, L is covalently attached to N'⁴, which means that when a' is 0, an atom in L forms a covalent bond with N'⁴ in the compound. Similarly, when a' is 1 and b' is 0, L is covalently attached to N'¹ ; when b' is 1 , and c' is 0, L is covalently attached to N'²; when c' is 1 , L is covalently attached to N'³; when a is 0, L is covalently attached to N⁴; when a is 1 and b is 0, L is covalently attached to N¹ ; when b is 1 , and c is 0, L is covalently attached to N²; and when c is 1 , L is covalently attached to N³.

The terms "linker," "Ι_'," "L," "L¹ ," and "L ," as used herein, refer to a covalent linkage or connection between two or more components in a compound (e.g., two cyclic heptapeptides and one or more monosaccharide or oligosaccharide moieties in a compound described herein). In some embodiments, a compound described herein may contain a linker that has a trivalent structure (e.g., a trivalent linker). A trivalent linker has three arms, in which each arm is conjugated to a component of the compound (e.g., a first arm conjugated to a first cyclic heptapeptide, a second arm conjugated to a second heptapeptide, and a third arm conjugated to one or more monosaccharide or oligosaccharide moieties). In some embodiments, a compound described herein may contain two or three linkers (e.g., a compound of formula (XIII) or (XIV)), in which each linker has a divalent structure (e.g., a divalent linker). For example, the first linker may connect the first and second cyclic heptapeptides (e.g., L in the compound of formula (XIII)), the second linker may connect a first monosaccharide or oligosaccharide moiety and a peptide (e.g., a peptide including a 1 -5 amino acid residue(s)) attached to the first cyclic heptapeptide (e.g., L¹ in the compound of formula (XIII)), and a third linker may connect a second monosaccharide or oligosaccharide moiety and a peptide (e.g., a peptide including a 1 -5 amino acid residue(s)) attached to the second cyclic heptapeptide (e.g., L'¹ in the compound of formula (XIII)).

Molecules that may be used as linkers include at least two functional groups, which may be the same or different, e.g., two carboxylic acid groups, two amine groups, two sulfonic acid groups, a carboxylic acid group and an amine group, or a carboxy group and a sulfonic acid group. The first functional group may form a covalent linkage with a first component in the compound and the second functional group may form a covalent linkage with the second component in the compound. In some embodiments of a trivalent linker, two arms of a linker may contain two dicarboxylic acids, in which the first carboxylic acid may form a covalent linkage with the first cyclic heptapeptide in the compound and the second carboxylic acid may form a covalent linkage with the second cyclic heptapeptide in the compound, and the third arm of the linker may for a covalent linkage (e.g., a C bond) with one or more monosaccharide or oligosaccharide moieties in the compound. In some embodiments of a divalent linker, the divalent linker may contain two carboxylic acids, in which the first carboxylic acid may form a covalent linkage with one component (e.g., a first cyclic heptapeptide) in the compound and the second carboxylic acid may form a covalent linkage with another component (e.g., a second cyclic heptapeptide) in the compound.

Examples of dicarboxylic acids are described further herein. In some embodiments, a molecule containing one or more sulfonic acid groups may be used as a linker, in which the sulfonic acid group may form a sulfonamide linkage with a component in the compound. In some embodiments, a molecule containing one or more isocyanate groups may be used as a linker, in which the isocyanate group may form a urea linkage with a component in the compound. In some embodiments, a molecule containing one or more haloalkyl groups may be used as a linker, in which the haloalkyl group may form a covalent linkage, e.g., C-N and C-0 linkages, with a component in the compound.

In some embodiments, a linker provides space, rigidity, and/or flexibility between the two or more components. In some embodiments, a linker may be a bond, e.g., a covalent bond. The term "bond" refers to a chemical bond, e.g., an amide bond, a disulfide bond, a C-0 bond, a N-N bond, or any kind of bond created from a chemical reaction, e.g., chemical conjugation. In some embodiments, a linker includes no more than 250 atoms. In some embodiments, a linker includes no more than 250 non- hydrogen atoms. In some embodiments, the backbone of a linker includes no more than 250 atoms. The "backbone" of a linker refers to the atoms in the linker that together form the shortest path from one part of a compound to another part of the compound (e.g., the shortest path linking a first cyclic heptapeptide and a second cyclic heptapeptide). The atoms in the backbone of the linker are directly involved in linking one part of a compound to another part of the compound (e.g., linking a first cyclic heptapeptide and a second cyclic heptapeptide). For examples, hydrogen atoms attached to carbons in the backbone of the linker are not considered as directly involved in linking one part of the compound to another part of the compound. In some embodiments, a linker may comprise a synthetic group derived from, e.g., a synthetic polymer (e.g., a polyethylene glycol (PEG) polymer). In some embodiments, a linker may comprise one or more amino acid residues, such as D- or L-amino acid residues. In some embodiments, a linker may be a residue of an amino acid sequence (e.g., a 1 -25 amino acid, 1 -10 amino acid, 1 -9 amino acid, 1 -8 amino acid, 1 -7 amino acid, 1 -6 amino acid, 1 -5 amino acid, 1 -4 amino acid, 1 -3 amino acid, 1 -2 amino acid, or 1 amino acid sequence). In some embodiments, a linker may comprise one or more, e.g., 1 -100, 1 -50, 1 -25, 1 -10, 1 -5, or 1 -3, optionally substituted alkylene, optionally substituted heteroalkylene (e.g., a PEG unit), optionally substituted alkenylene, optionally substituted heteroalkenylene, optionally substituted alkynylene, optionally substituted heteroalkynylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene, optionally substituted cycloalkenylene, optionally substituted heterocycloalkenylene, optionally substituted cycloalkynylene, optionally substituted

heterocycloalkynylene, optionally substituted arylene, optionally substituted heteroarylene (e.g., pyridine), O, S, NR' (Rⁱ is H, optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted alkenyl, optionally substituted heteroalkenyl, optionally substituted alkynyl, optionally substituted heteroalkynyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted cycloalkenyl, optionally substituted heterocycloalkenyl, optionally substituted cycloalkynyl, optionally substituted heterocycloalkynyl, optionally substituted aryl, or optionally substituted heteroaryl), P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino. For example, a linker may comprise one or more optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene (e.g., a PEG unit), optionally substituted C2-C20 alkenylene (e.g., C2 alkenylene), optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20

heteroalkynylene, optionally substituted C3-C20 cycloalkylene (e.g., cyclopropylene, cyclobutylene), optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene (e.g., C6 arylene), optionally substituted C2-C15 heteroarylene (e.g., imidazole, pyridine), O, S, NR' (R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2- C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl), P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino.

The term "lipophilic moiety," as used herein, refers to a portion, substituent, or functional group of a compound that is, in general, hydrophobic and non-polar. A moiety is lipophilic if it has a hydrophobicity determined using a cLogP value of greater than 0, such as about 0.25 or greater, about 0.5 or greater, about 1 or greater, about 2 or greater, 0.25-5, 0.5-4 or 2-3. As used herein, the term "cLogP" refers to the calculated partition coefficient of a molecule or portion of a molecule. The partition coefficient is the ratio of concentrations of a compound in a mixture of two immiscible phases at equilibrium (e.g., octanol and water) and measures the hydrophobicity or hydrophilicity of a compound. A variety of methods are available in the art for determining cLogP. For example, in some embodiments, cLog P can be determined using quantitative structure-property relationship algorithims known in the art (e.g. , using fragment based prediction methods that predict the logP of a compound by determining the sum of its non-overlapping molecular fragments). Several algorithims for calculating cLog P are known in the art including those used by molecular editing software such as CH EMDRAW® Pro, Version 12.0.2.1092

(Camrbridgesoft, Cambridge, MA) and MARVINSKETCH® (ChemAxon, Budapest, Hungary). A moiety is considered lipophilic if it has a cLogP value described above in at least one of the above methods. A lipophilic moiety having the stated cLogP value will be considered lipophilic, even though it may have a positive charge or a polar substituent.

In some embodiments, a lipophilic moiety contains entirely hydrocarbons. In some embodiments, a lipophilic moiety may contain one or more, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, heteroatoms independently selected from N, O, and S (e.g. , an indolyl), or one or more, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, halo groups, which, due to the structure of the moiety and/or small differences in electronegativity between the heteroatoms or halo groups and the hydrocarbons, do not induce significant chemical polarity into the lipophilic moiety. Thus, in some embodiments, a lipophilic moiety having, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, heteroatoms and/or, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, halo atoms may still be considered non-polar. In some embodiments, a lipophilic moiety may be optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl , or optionally substituted heteroaryl, or halo forms thereof, wherein the optional substituents are also lipophilic (such as alkyl, alkenyl , alkynyl, heteroalkyl, heteroalkenyl, heteroalkynyl, aryl , or heteroaryl) or are not lipophilic but do not change the overall lipophilic character of the moiety, i.e. , the moiety has a cLogP value of greater than 0. For example, octanol contains a polar group, OH, but is still a lipophilic moiety. In some embodiments, a lipophilic moiety may be benzyl, isobutyl, sec-butyl, isopropyl, n-propyl, methyl, biphenylmethyl, n-octyl , or substituted indolyl (e.g. , alkyl substituted indolyl). In some embodiments, a lipophilic moiety may be the side chain of a hydrophobic amino acid residue, e.g ., leucine, isoleucine, alanine, phenylalanine, valine, and proline, or groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl , and pyrrolidinyl. In some embodiments, lipophilic moieties of the compounds described herein may interact with the hydrophobic portions of lipid A (e.g. , fatty acid side chains of lipid A) when the compounds bind to the membrane of bacterial cells (e.g. , Gram-negative bacterial cells). Due to its position on the cyclic heptapeptide, one or more of R¹ , R¹², R¹⁵, R'¹ , R'¹², and R'¹⁵ may be a lipophilic moiety.

The term "positively charged moiety," as used herein, refers to a portion , substituent, or functional group of a compound that contains at least one positive charge. In some embodiments, a positively charged moiety contains one or more (e.g., 1 -4, 1 -3, 1 , 2, 3, or 4) heteroatoms independently selected from N, O, and S, for example. In some embodiments, a positively charged moiety may possess a pH- dependent positive charge, e.g. , the moiety becomes a positively charged moiety at physiological pH (e.g. , pH 7), such as -NH₃ ⁺, -(CH₂)₄NH2, -(CH₂)₃NH2, -(CH₂)₂NH2, -CH2NH2, -(CH₂)4N(CH₃)2,

-(CH2)₃N(CH₃)2, -(CH₂)₂N(CH₃)2, -CH₂N(CH₃)2, -(CH₂)4NH(CH₃), -(CH2)₃NH(CH₃),-(CH₂)2NH(CH₃), and -CH2NH(CH₃). In some embodiments, a positively charged moiety may be optionally substituted alkamino, optionally substituted heteroalkyl (e.g., optionally substituted heteroalkyl containing 1 -3 nitrogens; -(CH2)4-guanidinium, -(CH2)3-guanidinium, -(CH2)2-guanidinium, -CH2-guanidinium), optionally substituted heterocycloalkyl (e.g., optionally substituted heterocycloalkyl containing 1 -3 nitrogens), or optionally substituted heteroaryl (e.g., optionally substituted heteroaryl containing 1 -3

nitrogens; -(CH2)4-imidazole, -(CH2)3-imidazole, -(CH2)2-imidazole, -CH2-imidazole). In some

embodiments, a positively charged moiety may be pH independent such

as -CH₂N(CH₃)3⁺, -(CH2)₂N(CH₃)3⁺, -(CH2)₃N(CH₃)3⁺, or -(CH₂)4N(CH₃)3⁺. Thus, substituents may transform an otherwise lipophilic moiety such as optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, or optionally substituted heteroaryl, or halo forms thereof, to a positively charged moiety with the addition of a substituent that imparts a positive charge or a pH dependent positive charge, such as guanidinyl, -NH3⁺, -NH2, -NH(CH3), -N(CH3)2, and/or -N(CH3)3⁺. In some embodiments, a positively charged moiety may be the side chain of an amino acid residue (e.g., a natural or non-natural amino acid residue, such as a D- or L-amino acid residue, that is positively charged at physiological pH (e.g., pH 7), such as the side chain of a basic amino acid residue (e.g., arginine, lysine, histidine, ornithine, diaminobuteric acid, or diaminopropionic acid). In some embodiments, positively charged moieties of the compounds described herein interact with the negatively charged portions of lipid A (e.g., phosphates of lipid A) when the compounds bind to the membrane of bacterial cells (e.g., Gram-negative bacterial cells). Due to its position on the cyclic heptapeptide, one or more of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ may be a positively charged moiety.

The term "polar moiety," as used herein, refers to a portion, substituent, or functional group of a compound that has a chemical polarity induced by atoms with different electronegativity. The polarity of a polar moiety is dependent on the electronegativity between atoms within the moiety and the asymmetry of the structure of the moiety. In some embodiments, a polar moiety contains one or more (e.g., 1 -4, 1 -3, 1 , 2, 3, or 4) heteroatoms independently selected from N, O, and S, which may induce chemical polarity in the moiety by having different electronegativity from carbon and hydrogen. In general, a polar moiety interacts with other polar or charged molecules. In some embodiments, a polar moiety may be optionally substituted alkamino, optionally substituted heteroalkyl (e.g., N- and/or O-containing

heteroalkyl; -(CH2)4-carboxylic acid, -(CH2)3-carboxylic acid, -(CH2)2-carboxylic acid, -CH2-carboxylic acid), optionally substituted heterocycloalkyl (e.g., N- and/or O-containing heterocycloalkyl), or optionally substituted heteroaryl (e.g., N- and/or O-containing heteroaryl). In some embodiments, a polar moiety may -CH(CH₃)OH, -CH2OH, -(CH₂)₂CONH2, -CH2CONH2, -CH2COOH, or -(CH₂)₂COOH. Thus, substituents may transform an otherwise lipophilic moiety such optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, or optionally substituted heteroaryl, or halo forms thereof, to a polar moiety with the addition of a substituent that imparts polarity, such as -OH, -COOH, -COOR, or -CONR2, in which R is H or C1 -C4 alkyl. In some embodiments, a polar moiety may be the side chain or a polar or charged amino acid residue (e.g., threonine, serine, glutamine, asparagine, arginine, lysine histidine, aspartic acid, and glutamic acid). In some embodiments, a polar moiety is the side chain of threonine. In some embodiments, polar moieties of the compounds described herein interact with the negatively charged portions of lipid A (e.g., phosphates of lipid A) when the compounds bind to the membrane of bacterial cells (e.g., Gram-negative bacterial cells). Due to its position on the cyclic heptapeptide, one or more of R¹ , R¹², R¹⁵, R'¹ , R'¹², and R'¹⁵ may be a polar moiety.

The term "polymyxin core," as used herein means a cyclic heptapeptide having the structure:

wherein Q¹ , Q² and Q³ are as follows:

attachment of the polymyxin core to the remainder of the compounds disclosed herein, including the second polymyxin core, the linker, and E groups of the compounds disclosed herein.

The terms "alkyl," "alkenyl," and "alkynyl," as used herein, include straight-chain and branched- chain monovalent substituents, as well as combinations of these, containing only C and H when unsubstituted. When the alkyl group includes at least one carbon-carbon double bond or carbon-carbon triple bond, the alkyl group can be referred to as an "alkenyl" or "alkynyl" group respectively. The monovalency of an alkyl, alkenyl, or alkynyl group does not include the optional substituents on the alkyl, alkenyl, or alkynyl group. For example, if an alkyl, alkenyl, or alkynyl group is attached to a compound, monovalency of the alkyl, alkenyl, or alkynyl group refers to its attachment to the compound and does not include any additional substituents that may be present on the alkyl, alkenyl, or alkynyl group. In some embodiments, the alkyl or heteroalkyl group may contain, e.g., 1 -20. 1 -18, 1 -16, 1 -14, 1 -12, 1 -10, 1 -8, 1 - 6, 1 -4, or 1 -2 carbon atoms (e.g., C1 -C20, C1 -C18, C1 -C16, C1 -C14, C1 -C12, C1 -C10, C1 -C8, C1 -C6, C1 -C4, or C1 -C2). In some embodiments, the alkenyl, heteroalkenyl, alkynyl, or heteroalkynyl group may contain, e.g., 2-20, 2-18, 2-16, 2-14, 2-12, 2-10, 2-8, 2-6, or 2-4 carbon atoms (e.g., C2-C20, C2-C18, C2-C16, C2-C14, C2-C12, C2-C10, C2-C8, C2-C6, or C2-C4). Examples include, but are not limited to, methyl, ethyl, isobutyl, sec-butyl, tert-butyl, 2-propenyl, and 3-butynyl.

The term "cycloalkyl," as used herein, represents a monovalent saturated or unsaturated non- aromatic cyclic alkyl group. A cycloalkyl may have, e.g., three to twenty carbons (e.g., a C3-C7, C3-C8, C3-C9, C3-C10, C3-C1 1 , C3-C12, C3-C14, C3-C16, C3-C18, or C3-C20 cycloalkyi). Examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. When the cycloalkyi group includes at least one carbon-carbon double bond, the cycloalkyi group can be referred to as a "cycloalkenyl" group. A cycloalkenyl may have, e.g., four to twenty carbons (e.g., a C4- C7, C4-C8, C4-C9, C4-C10, C4-C1 1 , C4-C12, C4-C14, C4-C16, C4-C18, or C4-C20 cycloalkenyl).

Exemplary cycloalkenyl groups include, but are not limited to, cyclopentenyl, cyclohexenyl, and cycloheptenyl. When the cycloalkyi group includes at least one carbon-carbon triple bond, the cycloalkyi group can be referred to as a "cycloalkynyl" group. A cycloalkynyl may have, e.g., eight to twenty carbons (e.g., a C8-C9, C8-C10, C8-C1 1 , C8-C12, C8-C14, C8-C16, C8-C18, or C8-C20 cycloalkynyl). The term "cycloalkyi" also includes a cyclic compound having a bridged multicyclic structure in which one or more carbons bridges two non-adjacent members of a monocyclic ring, e.g., bicyclo[2.2.1 .]heptyl and adamantane. The term "cycloalkyi" also includes bicyclic, tricyclic, and tetracyclic fused ring structures, e.g., decalin and spiro cyclic compounds.

The term "aryl," as used herein, refers to any monocyclic or fused ring bicyclic or tricyclic system which has the characteristics of aromaticity in terms of electron distribution throughout the ring system, e.g., phenyl, naphthyl, or phenanthrene. In some embodiments, a ring system contains 5-15 ring member atoms or 5-10 ring member atoms. An aryl group may have, e.g., five to fifteen carbons (e.g., a C5-C6, C5-C7, C5-C8, C5-C9, C5-C10, C5-C1 1 , C5-C12, C5-C13, C5-C14, or C5-C15 aryl). The term

"heteroaryl" also refers to such monocyclic or fused bicyclic ring systems containing one or more, e.g., 1 - 4, 1 -3, 1 , 2, 3, or 4, heteroatoms selected from O, S and N. A heteroaryl group may have, e.g., two to fifteen carbons (e.g., a C2-C3, C2-C4, C2-C5, C2-C6, C2-C7, C2-C8, C2-C9. C2-C10, C2-C1 1 , C2-C12, C2-C13, C2-C14, or C2-C15 heteroaryl). The inclusion of a heteroatom permits inclusion of 5-membered rings to be considered aromatic as well as 6-membered rings. Thus, typical heteroaryl systems include, e.g., pyridyl, pyrimidyl, indolyl, benzimidazolyl, benzotriazolyl, isoquinolyl, quinolyl, benzothiazolyl, benzofuranyl, thienyl, furyl, pyrrolyl, thiazolyl, oxazolyl, isoxazolyl, benzoxazolyl, benzoisoxazolyl, and imidazolyl. Because tautomers are possible, a group such as phthalimido is also considered heteroaryl. In some embodiments, the aryl or heteroaryl group is a 5- or 6-membered aromatic rings system optionally containing 1 -2 nitrogen atoms. In some embodiments, the aryl or heteroaryl group is an optionally substituted phenyl, pyridyl, indolyl, pyrimidyl, pyridazinyl, benzothiazolyl, benzimidazolyl, pyrazolyl, imidazolyl, isoxazolyl, thiazolyl, or imidazopyridinyl. In some embodiments, the aryl group is phenyl. In some embodiments, an aryl group may be optionally substituted with a substituent such an aryl substituent, e.g., biphenyl.

The term "alkaryl," refers to an aryl group that is connected to an alkylene, alkenylene, or alkynylene group. In general, if a compound is attached to an alkaryl group, the alkylene, alkenylene, or alkynylene portion of the alkaryl is attached to the compound. In some embodiments, an alkaryl is C6- C35 alkaryl (e.g., C6-C16, C6-C14, C6-C12, C6-C10, C6-C9, C6-C8, C7, or C6 alkaryl), in which the number of carbons indicates the total number of carbons in both the aryl portion and the alkylene, alkenylene, or alkynylene portion of the alkaryl. Examples of alkaryls include, but are not limited to, (C1 - C8)alkylene(C6-C12)aryl, (C2-C8)alkenylene(C6-C12)aryl, or (C2-C8)alkynylene(C6-C12)aryl. In some embodiments, an alkaryl is benzyl. In a heteroalkaryl, one or more heteroatoms selected from N, O, and S may be present in the alkylene, alkenylene, or alkynylene portion of the alkaryl group and/or may be present in the aryl portion of the alkaryl group. In an optionally substituted alkaryl, the substituent may be present on the alkylene, alkenylene, or alkynylene portion of the alkaryl group and/or may be present on the aryl portion of the alkaryl group.

The term "amino," as used herein, represents -N(R^X)2 or -N⁺(R^X)3, where each R^x is,

independently, H, alkyl, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, or two R^x combine to form a

heterocycloalkyl. In some embodiment, the amino group is -NH2.

The term "alkamino," as used herein, refers to an amino group, described herein, that is attached to an alkylene (e.g., C1 -C5 alkylene), alkenylene (e.g., C2-C5 alkenylene), or alkynylene group (e.g., C2- C5 alkenylene). In general, if a compound is attached to an alkamino group, the alkylene, alkenylene, or alkynylene portion of the alkamino is attached to the compound. The amino portion of an alkamino refers to -N(R^X)2 or -N⁺(R^X)3, where each R^x is, independently, H, alkyl, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, or two R^x combine to form a heterocycloalkyl. In some embodiment, the amino portion of an alkamino is -NH2. An example of an alkamino group is C1 -C5 alkamino, e.g., C2 alkamino (e.g., CH2CH2NH2 or CH2CH2N(CH3)2). In a heteroalkamino group, one or more, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, heteroatoms selected from N, O, and S may be present in the alkylene, alkenylene, or alkynylene portion of the heteroalkamino group. In some embodiments, an alkamino group may be optionally substituted. In a substituted alkamino group, the substituent may be present on the alkylene, alkenylene, or alkynylene portion of the alkamino group and/or may be present on the amino portion of the alkamino group.

The term "alkamide," as used herein, refers to an amide group that is attached to an alkylene (e.g., C1 -C5 alkylene), alkenylene (e.g., C2-C5 alkenylene), or alkynylene (e.g., C2-C5 alkenylene) group. In general, if a compound is attached to an alkamide group, the alkylene, alkenylene, or alkynylene portion of the alkamide is attached to the compound. The amide portion of an alkamide refers to -C(0)-N(R^X)2, where each R^x is, independently, H, alkyl, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, or two R^x combine to form a heterocycloalkyl. In some embodiment, the amide portion of an alkamide is -C(0)NH2. An alkamide group may be -(CH2)2-C(0)NH2 or -CH2-C(0)NH2. In a heteroalkamide group, one or more, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, heteroatoms selected from N, O, and S may be present in the alkylene, alkenylene, or alkynylene portion of the heteroalkamide group. In some embodiments, an alkamide group may be optionally substituted. In a substituted alkamide group, the substituent may be present on the alkylene, alkenylene, or alkynylene portion of the alkamide group and/or may be present on the amide portion of the alkamide group.

The terms "alkylene," "alkenylene," and "alkynylene," as used herein, refer to divalent groups having a specified size. In some embodiments, an alkylene may contain, e.g., 1 -20, 1 -18, 1 -16, 1 -14, 1 - 12, 1 -10, 1 -8, 1 -6, 1 -4, or 1 -2 carbon atoms (e.g., C1 -C20, C1 -C18, C1 -C1 6, C1 -C14, C1 -C12, C1 -C1 0, C1 -C8, C1 -C6, C1 -C4, or C1 -C2). In some embodiments, an alkenylene or alkynylene may contain, e.g., 2-20, 2-18, 2-16, 2-14, 2-12, 2-10, 2-8, 2-6, or 2-4 carbon atoms (e.g., C2-C20, C2-C18, C2-C16, C2- C14, C2-C12, C2-C10, C2-C8, C2-C6, or C2-C4). Alkylene, alkenylene, and/or alkynylene includes straight-chain and branched-chain forms, as well as combinations of these. The divalency of an alkylene, alkenylene, or alkynylene group does not include the optional substituents on the alkylene, alkenylene, or alkynylene group. For example, two cyclic heptapeptides may be attached to each other by way of a linker that includes alkylene, alkenylene, and/or alkynylene, or combinations thereof. Each of the alkylene, alkenylene, and/or alkynylene groups in the linker is considered divalent with respect to the two attachments on either end of alkylene, alkenylene, and/or alkynylene group. For example, if a linker includes -(optionally substituted alkylene)-(optionally substituted alkenylene)-(optionally substituted alkylene)-, the alkenylene is considered divalent with respect to its attachments to the two alkylenes at the ends of the linker. The optional substituents on the alkenylene are not included in the divalency of the alkenylene. The divalent nature of an alkylene, alkenylene, or alkynylene group (e.g., an alkylene, alkenylene, or alkynylene group in a linker) refers to both of the ends of the group and does not include optional substituents that may be present in an alkylene, alkenylene, or alkynylene group. Because they are divalent, they can link together multiple (e.g., two) parts of a compound, e.g., a first cyclic heptapeptide and a second cyclic heptapeptide. Alkylene, alkenylene, and/or alkynylene groups can be substituted by the groups typically suitable as substituents for alkyl, alkenyl and alkynyl groups as set forth herein. For example, C=0 is a C1 alkylene that is substituted by an oxo (=0). For

example, -HCR-C≡C- may be considered as an optionally substituted alkynylene and is considered a divalent group even though it has an optional substituent, R. Heteroalkylene, heteroalkenylene, and/or heteroalkynylene groups refer to alkylene, alkenylene, and/or alkynylene groups including one or more, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, heteroatoms, e.g., N, O, and S. For example, a polyethylene glycol (PEG) polymer or a PEG unit -(CH2)2-0- in a PEG polymer is considered a heteroalkylene containing one or more oxygen atoms.

The term "cycloalkylene," as used herein, refers to a divalent cyclic group linking together two parts of a compound. For example, one carbon within the cycloalkylene group may be linked to one part of the compound, while another carbon within the cycloalkylene group may be linked to another part of the compound. A cycloalkylene group may include saturated or unsaturated non-aromatic cyclic groups. A cycloalkylene may have, e.g., three to twenty carbons in the cyclic portion of the cycloalkylene (e.g., a C3-C7, C3-C8, C3-C9, C3-C10, C3-C1 1 , C3-C12, C3-C14, C3-C16, C3-C18, or C3-C20 cycloalkylene). When the cycloalkylene group includes at least one carbon-carbon double bond, the cycloalkylene group can be referred to as a "cycloalkenylene" group. A cycloalkenylene may have, e.g., four to twenty carbons in the cyclic portion of the cycloalkenylene (e.g., a C4-C7, C4-C8, C4-C9. C4-C10, C4-C1 1 , C4- C12, C4-C14, C4-C16, C4-C18, or C4-C20 cycloalkenylene). When the cycloalkylene group includes at least one carbon-carbon triple bond, the cycloalkylene group can be referred to as a "cycloalkynylene" group. A cycloalkynylene may have, e.g., four to twenty carbons in the cyclic portion of the

cycloalkynylene (e.g., a C4-C7, C4-C8, C4-C9. C4-C1 0, C4-C1 1 , C4-C12, C4-C14, C4-C16, C4-C18, or C8-C20 cycloalkynylene). A cycloalkylene group can be substituted by the groups typically suitable as substituents for alkyl, alkenyl and alkynyl groups as set forth herein. Heterocycloalkylene refers to a cycloalkylene group including one or more, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, heteroatoms, e.g., N, O, and S. Examples of cycloalkylenes include, but are not limited to, cyclopropylene and cyclobutylene. A tetrahydrofuran may be considered as a heterocycloalkylene.

The term "arylene," as used herein, refers to a multivalent (e.g., divalent or trivalent) aryl group linking together multiple (e.g., two or three) parts of a compound. For example, one carbon within the arylene group may be linked to one part of the compound, while another carbon within the arylene group may be linked to another part of the compound. An arylene may have, e.g., five to fifteen carbons in the aryl portion of the arylene (e.g., a C5-C6, C5-C7, C5-C8, C5-C9. C5-C10, C5-C1 1 , C5-C12, C5-C13, C5- C14, or C5-C15 arylene). An arylene group can be substituted by the groups typically suitable as substituents for alkyl, alkenyl and alkynyl groups as set forth herein. Heteroarylene refers to an aromatic group including one or more, e.g., 1 -4, 1 -3, 1 , 2, 3, or 4, heteroatoms, e.g., N, O, and S. A heteroarylene group may have, e.g., two to fifteen carbons (e.g., a C2-C3, C2-C4, C2-C5, C2-C6, C2-C7, C2-C8, C2- C9. C2-C10, C2-C1 1 , C2-C12, C2-C13, C2-C14, or C2-C15 heteroarylene).

The term "optionally substituted," as used herein, refers to having 0, 1 , or more substituents, such as 0-25, 0-20, 0-10 or 0-5 substituents. Substituents include, but are not limited to, alkyl, alkenyl, alkynyl, aryl, alkaryl, acyl, heteroaryl, heteroalkyl, heteroalkenyl, heteroalkynyl, heteroalkaryl, halogen, oxo, cyano, nitro, amino, alkamino, hydroxy, alkoxy, alkanoyl, carbonyl, carbamoyl, guanidinyl, ureido, amidinyl, any of the groups or moieties described above, and hetero versions of any of the groups or moieties described above. Substituents include, but are not limited to, F, CI, methyl, phenyl, benzyl, OR, NR₂, SR, SOR, S0₂R, OCOR, NRCOR, NRCONR2, NRCOOR, OCONR2, RCO, COOR, alkyl-OOCR, SO3R, CONR2, SO2NR2, NRSO2NR2, CN, CF₃, OCF3, SiR₃, and NO2, wherein each R is, independently, H, alkyl, alkenyl, aryl, heteroalkyl, heteroalkenyl, or heteroaryl, and wherein two of the optional substituents on the same or adjacent atoms can be joined to form a fused, optionally substituted aromatic or nonaromatic, saturated or unsaturated ring which contains 3-8 members, or two of the optional substituents on the same atom can be joined to form an optionally substituted aromatic or nonaromatic, saturated or unsaturated ring which contains 3-8 members.

An optionally substituted group or moiety refers to a group or moiety (e.g., any one of the groups or moieties described above) in which one of the atoms (e.g., a hydrogen atom) is optionally replaced with another substituent. For example, an optionally substituted alkyl may be an optionally substituted methyl, in which a hydrogen atom of the methyl group is replaced by, e.g., OH. As another example, a substituent on a heteroalkyl or its divalent counterpart, heteroalkylene, may replace a hydrogen on a carbon or a hydrogen on a heteroatom such as N. For example, the hydrogen atom in the

group -R-NH-R- may be substituted with an alkamide substituent, e.g., -R-N[(CH₂C(0)N(CH₃)2]-R.

Generally, an optional substituent is a noninterfering substituent. A "noninterfering substituent" refers to a substituent that leaves the ability of the compounds described herein (e.g., compounds of any one of formulas (l)-(XXXXX)) to either bind to lipopolysaccharides (LPS) or to kill or inhibit the growth of Gram- negative bacteria qualitatively intact. Thus, in some embodiments, the substituent may alter the degree of such activity. However, as long as the compound retains the ability to bind to LPS and/or to kill or inhibit the growth of Gram-negative bacteria, the substituent will be classified as "noninterfering." In some aspects, a noninterfering substituent leaves the ability of a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) to kill or inhibit the growth of Gram-negative bacteria qualitatively intact as determined by measuring the minimum inhibitory concentration (MIC) against at least one Gram- negative bacteria as known in the art, wherein the MIC is 32 μg/mL or less. In some aspects, a noninterfering substituent leaves the ability of a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) to bind to lipopolysaccharides (LPS) from the cell membrane of Gram-negative bacteria qualitatively intact, as determined by an LPS binding assay (e.g., see Example 103), wherein the compound shows a value of about 1 0% or greater displacement of a fluorogenic substrate at 250 μΜ of the compound.

The term "hetero," when used to describe a chemical group or moiety, refers to having at least one heteroatom that is not a carbon or a hydrogen, e.g., N, O, and S. Any one of the groups or moieties described above may be referred to as hetero if it contains at least one heteroatom. For example, a heterocycloalkyi, heterocycloalkenyl, or heterocycloalkynyl group refers to a cycloalkyi, cycloalkenyl, or cycloalkynyl group that has one or more heteroatoms independently selected from, e.g., N, O, and S. An example of a heterocycloalkenyl group is a maleimido. For example, a heteroaryl group refers to an aromatic group that has one or more heteroatoms independently selected from, e.g., N, O, and S. One or more heteroatoms may also be included in a substituent that replaced a hydrogen atom in a group or moiety as described herein. For example, in an optionally substituted heteroaryl group, if one of the hydrogen atoms in the heteroaryl group is replaced with a substituent (e.g., methyl), the substituent may also contain one or more heteroatoms (e.g., methanol).

The term "acyl," as used herein, refers to a group having the structure:

, wherein R^z is an optionally substituted alkyl, alkenyl, alkynyl, cycloalkyi, cycloalkenyl, cycloalkynyl, aryl, alkaryl, alkamino, heteroalkyl, heteroalkenyl, heteroalkynyl, heterocycloalkyi, heterocycloalkenyl,

heterocycloalkynyl, heteroaryl, heteroalkaryl, or heteroalkamino.

The term "halo" or "halogen," as used herein, refers to any halogen atom, e.g., F, CI, Br, or I. Any one of the groups or moieties described herein may be referred to as a "halo moiety" if it contains at least one halogen atom, such as haloalkyl.

The term "hydroxyl," as used herein, represents an -OH group.

The term "oxo," as used herein, refers to a substituent having the structure =0, where there is a double bond between an atom and an oxygen atom.

O

The term "carbonyl," as used herein, refers to a group having the structure

s

The term "thiocarbonyl," as used herein, refers to a group having the structure: ^ ^

O

i I I l-o-^p I -c

The term "phosphate," as used herein, represents the group having the structure: O^"

O

¾ I I

|— ^p-o-

The term "phosphoryl," as used herein, represents the group having the structure: OR

O

4 I I <,

i-o-^p-o-

R

The term "sulfonyl," as used herein, represents the group having the structure V NR

The term "imino," as used herein, represents the group having the structure: ^ ^ , wherein R is an optional substituent.

The term "AV-protecting group," as used herein, represents those groups intended to protect an amino group against undesirable reactions during synthetic procedures. Commonly used AV-protecting groups are disclosed in Greene, "Protective Groups in Organic Synthesis," 5th Edition (John Wiley & Sons, New York, 2014), which is incorporated herein by reference. AV-protecting groups include, e.g., acyl, aryloyl, and carbamyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, a-chlorobutyryl, benzoyl, carboxybenzyl (CBz), 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and chiral auxiliaries such as protected or unprotected D, L or D, L-amino acid residues such as alanine, leucine, phenylalanine;

sulfonyl-containing groups such as benzenesulfonyl and p-toluenesulfonyl ; carbamate forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p- nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4- dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyl oxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl,

3,4,5-trimethoxybenzyloxycarbonyl, 1 -(p-biphenylyl)-l -methylethoxycarbonyl, α,α-dimethyl- 3,5-dimethoxybenzyloxycarbonyl, benzhydryloxy carbonyl, t-butyloxycarbonyl (BOC),

diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxy carbonyl, fluorenyl-9-methoxycarbonyl (Fmoc), cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, and phenylthiocarbonyl; alkaryl groups such as benzyl, triphenylmethyl, and benzyloxymethyl ; and silyl groups such as trimethylsilyl.

The term "amino acid," as used herein, means naturally occurring amino acids and non-naturally occurring amino acids.

The term "naturally occurring amino acids," as used herein, means amino acids including Ala,

Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, lie, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val.

The term "non-naturally occurring amino acid," as used herein, means an alpha amino acid that is not naturally produced or found in a mammal. Examples of non-naturally occurring amino acids include D- amino acids; an amino acid having an acetylaminomethyl group attached to a sulfur atom of a cysteine; a pegylated amino acid; the omega amino acids of the formula NH2(CH2)_nCOOH where n is 2-6, neutral nonpolar amino acids, such as sarcosine, t-butyl alanine, t-butyl glycine, N-methyl isoleucine, and norleucine; oxymethionine; phenylglycine; citrulline; methionine sulfoxide; cysteic acid; ornithine;

diaminobutyric acid; 3-aminoalanine; 3-hydroxy-D-proline; 2,4-diaminobutyric acid; 2-aminopentanoic acid; 2-aminooctanoic acid, 2-carboxy piperazine; piperazine-2-carboxylic acid, 2-amino-4-phenylbutanoic acid; 3-(2-naphthyl)alanine, and hydroxyproline. Other amino acids are a-aminobutyric acid, a-amino-a- methylbutyrate, aminocyclopropane-carboxylate, aminoisobutyric acid, aminonorbornyl-carboxylate, L- cyclohexylalanine, cyclopentylalanine, L-N-methylleucine, L-N-methylmethionine, L-N-methylnorvaline, L- N-methylphenylalanine, L-N-methylproline, L-N-methylserine, L-N-methyltryptophan, D-ornithine, L-N- methylethylglycine, L-norleucine, a-methyl-aminoisobutyrate, a-methylcyclohexylalanine, D-a- methylalanine, D-a-methylarginine, D-a-methylasparagine, D-a-methylaspartate, D-a-methylcysteine, D- a-methylglutamine, D-a-methylhistidine, D-a-methylisoleucine, D-a-methylleucine, D-a-methyllysine, D-a- methylmethionine, D-a-methylornithine, D-a-methylphenylalanine, D-a-methylproline, D-a-methylserine, D-N-methylserine, D-a-methylthreonine, D-a-methyltryptophan, D-a-methyltyrosine, D-a-methylvaline, D- N-methylalanine, D-N-methylarginine, D-N-methylasparagine, D-N-methylaspartate, D-N-methylcysteine, D-N-methylglutamine, D-N-methylglutamate, D-N-methylhistidine, D-N-methylisoleucine, D-N- methylleucine, D-N-methyllysine, N-methylcyclohexylalanine, D-N-methylornithine, N-methylglycine, N- methylaminoisobutyrate, N-(1 -methylpropyl)glycine, N-(2-methylpropyl)glycine, D-N-methyltryptophan, D- N-methyltyrosine, D-N-methylvaline, γ-aminobutyric acid, L-t-butylglycine, L-ethylglycine, L- homophenylalanine, L-a-methylarginine, L-a-methylaspartate, L-a-methylcysteine, L-a-methylglutamine, L-a-methylhistidine, L-a-methylisoleucine, L-a-methylleucine, L-a-methylmethionine, L-a-methylnorvaline, L-a-methylphenylalanine, L-a-methylserine, L-a-methyltryptophan, L-a-methylvaline, N-(N-(2,2- diphenylethyl) carbamylmethylglycine, 1 -carboxy-1 -(2,2-diphenyl-ethylamino) cyclopropane, 4- hydroxyproline, ornithine, 2-aminobenzoyl (anthraniloyl), D-cyclohexylalanine, 4-phenyl-phenylalanine, L- citrulline, a-cyclohexylglycine, L-1 ,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, L-thiazolidine-4- carboxylic acid, L-homotyrosine, L-2-furylalanine, L-histidine (3-methyl), N-(3-guanidinopropyl)glycine, O- methyl-L-tyrosine, O-glycan-serine, meta-tyrosine, nor-tyrosine, L-N,N',N"-trimethyllysine, homolysine, norlysine, N-glycan asparagine, 7-hydroxy-1 ,2,3,4-tetrahydro-4-fluorophenylalanine, 4- methylphenylalanine, bis-(2-picolyl)amine, pentafluorophenylalanine, indoline-2-carboxylic acid, 2- aminobenzoic acid, 3-amino-2-naphthoic acid, asymmetric dimethylarginine, L-tetrahydroisoquinoline-1 - carboxylic acid, D-tetrahydroisoquinoline-1 -carboxylic acid, 1 -amino-cyclohexane acetic acid, D/L- allylglycine, 4-aminobenzoic acid, 1 -amino-cyclobutane carboxylic acid, 2 or 3 or 4-aminocyclohexane carboxylic acid, 1 -amino-1 -cyclopentane carboxylic acid, 1 -aminoindane-1 -carboxylic acid, 4-amino- pyrrolidine-2-carboxylic acid, 2-aminotetraline-2-carboxylic acid, azetidine-3-carboxylic acid, 4-benzyl- pyrrolidine-2-carboxylic acid, tert-butylglycine, b-(benzothiazolyl-2-yl)-alanine, b-cyclopropyl alanine, 5,5- dimethyl-1 ,3-thiazolidine-4-carboxylic acid, (2R,4S)4-hydroxypiperidine-2-carboxylic acid, (2S,4S) and (2S,4R)-4-(2-naphthylmethoxy)-pyrrolidine-2-carboxylic acid, (2S,4S) and (2S,4R)4-phenoxy-pyrrolidine- 2-carboxylic acid, (2R,5S)and(2S,5R)-5-phenyl-pyrrolidine-2-carboxylic acid, (2S,4S)-4-amino-1 -benzoyl- pyrrolidine-2-carboxylic acid, t-butylalanine, (2S,5R)-5-phenyl-pyrrolidine-2-carboxylic acid, 1 - aminomethyl-cyclohexane-acetic acid, 3,5-bis-(2-amino)ethoxy-benzoic acid, 3,5-diamino-benzoic acid, 2- methylamino-benzoic acid, N-methylanthranylic acid, L-N-methylalanine, L-N-methylarginine, L-N- methylasparagine, L-N-methylaspartic acid, L-N-methylcysteine, L-N-methylglutamine, L-N- methylglutamic acid, L-N-methylhistidine, L-N-methylisoleucine, L-N-methyllysine, L-N-methylnorleucine, L-N-methylornithine, L-N-methylthreonine, L-N-methyltyrosine, L-N-methylvaline, L-N-methyl-t- butylglycine, L-norvaline, a-methyl-Y-aminobutyrate, 4,4'-biphenylalanine, a-methylcylcopentylalanine, a- methyl-a-napthylalanine, a-methylpenicillamine, N-(4-aminobutyl)glycine, N-(2-aminoethyl)glycine, N-(3- aminopropyl)glycine, N-amino-a-methylbutyrate, a-napthylalanine, N-benzylglycine, N-(2- carbamylethyl)glycine, N-(carbamylmethyl)glycine, N-(2-carboxyethyl)glycine, N-(carboxymethyl)glycine, N-cyclobutylglycine, N-cyclodecylglycine, N-cycloheptylglycine, N-cyclohexylglycine, N-cyclodecylglycine, N-cylcododecylglycine, N-cyclooctylglycine, N-cyclopropylglycine, N-cycloundecylglycine, N-(2,2- diphenylethyl)glycine, N-(3,3-diphenylpropyl)glycine, N-(3-guanidinopropyl)glycine, N-(1 - hydroxyethyl)glycine, N-(hydroxyethyl))glycine, N-(imidazolylethyl))glycine, N-(3-indolylyethyl)glycine, N- methyl-Y-aminobutyrate, D-N-methylmethionine, N-methylcyclopentylalanine, D-N-methylphenylalanine, D-N-methylproline, D-N-methylthreonine, N-(1 -methylethyl)glycine, N-methyl-napthylalanine, N- methylpenicillamine, N-(p-hydroxyphenyl)glycine, N-(thiomethyl)glycine, penicillamine, L-a-methylalanine, L-a-methylasparagine, L-a-methyl-t-butylglycine, L-methylethylglycine, L-a-methylglutamate, L-a- methylhomophenylalanine, N-(2-methylthioethyl)glycine, L-a-methyllysine, L-a-methylnorleucine, L-a- methylornithine, L-a-methylproline, L-a-methylthreonine, L-a-methyltyrosine, L-N-methyl- homophenylalanine, N-(N-(3,3-diphenylpropyl) carbamylmethylglycine, L-pyroglutamic acid, D- pyroglutamic acid, O-methyl-L-serine, O-methyl-L-homoserine, 5-hydroxylysine, a-carboxyglutamate, phenylglycine, L-pipecolic acid (homoproline), L-homoleucine, L-lysine (dimethyl), L-2-naphthylalanine, L- dimethyldopa or L-dimethoxy-phenylalanine, L-3-pyridylalanine, L-histidine (benzoyloxymethyl), N- cycloheptylglycine, L-diphenylalanine, O-methyl-L-homotyrosine, L-p-homolysine, O-glycan-threoine, Ortho-tyrosine, L-N,N'-dimethyllysine, L-homoarginine, neotryptophan, 3-benzothienylalanine, isoquinoline-3-carboxylic acid, diaminopropionic acid, homocysteine, 3,4-dimethoxyphenylalanine, 4- chlorophenylalanine, L-1 ,2,3,4-tetrahydronorharman-3-carboxylic acid, adamantylalanine, symmetrical dimethylarginine, 3-carboxythiomorpholine, D-1 ,2,3,4-tetrahydronorharman-3-carboxylic acid, 3- aminobenzoic acid, 3-amino-1 -carboxymethyl-pyridin-2-one, 1 -amino-1 -cyclohexane carboxylic acid, 2- aminocyclopentane carboxylic acid, 1 -amino-1 -cyclopropane carboxylic acid, 2-aminoindane-2-carboxylic acid, 4-amino-tetrahydrothiopyran-4-carboxylic acid, azetidine-2-carboxylic acid, b-(benzothiazol-2-yl)- alanine, neopentylglycine, 2-carboxymethyl piperidine, b-cyclobutyl alanine, allylglycine, diaminopropionic acid, homo-cyclohexyl alanine, (2S,4R)- 4-hydroxypiperidine-2-carboxylic acid, octahydroindole-2- carboxylic acid, (2S,4R) and (2S,4R)-4-(2-naphthyl), pyrrolidine-2-carboxylic acid, nipecotic acid, (2S,4R)and (2S,4S)-4-(4-phenylbenzyl) pyrrolidine-2-carboxylic acid, (3S)-1 -pyrrolidine-3-carboxylic acid, (2S,4S)-4-tritylmercapto-pyrrolidine-2-carboxylic acid, (2S,4S)-4-mercaptoproline, t-butylglycine, N,N- bis(3-aminopropyl)glycine, 1 -amino-cyclohexane-1 -carboxylic acid, N-mercaptoethylglycine, and selenocysteine. In some embodiments, amino acid residues may be charged or polar. Charged amino acids include alanine, lysine, aspartic acid, or glutamic acid, or non-naturally occurring analogs thereof. Polar amino acids include glutamine, asparagine, histidine, serine, threonine, tyrosine, methionine, or tryptophan, or non-naturally occurring analogs thereof.

It is specifically contemplated that in some embodiments, a terminal amino group in the amino acid may be an amido group or a carbamate group.

The term "antibacterial agent," as used herein, refers to an agent that is used in addition to one or more of the compounds described herein (e.g., compounds of any one of formulas (l)-(XXXXX)) in methods of treating a bacterial infection (e.g., Gram-negative bacterial infection) and/or preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria. An antibacterial agent may be an agent that prevents the entrance of a bacteria (e.g., a Gram-negative bacteria) into a subject's cells, tissues, or organs, inhibits the growth of a bacteria (e.g., a Gram-negative bacteria) in a subject's cells, tissues, or organs, and/or kills a bacteria (e.g., a Gram-negative bacteria) that is inside a subject's cells, tissues, or organs. Examples of antibacterial agents are described in detail further herein. In some embodiments, an antibacterial agent used in addition to a compound described herein is linezolid or tedizolid (e.g. , tedizolid phosphate).

The term "bacterial infection," as used herein, refers to the invasion of a subject's cells, tissues, and/or organs by bacteria (e.g., Gram-negative bacteria), thus, causing an infection. In some

embodiments, the bacteria may grow, multiply, and/or produce toxins in the subject's cells, tissues, and/or organs. In some embodiments, a bacterial infection can be any situation in which the presence of a bacterial population(s) is latent within or damaging to a host body. Thus, a subject is "suffering" from a bacterial infection when a latent bacterial population is detectable in or on the subject's body, an excessive amount of a bacterial population is present in or on the subject's body, or when the presence of a bacterial population(s) is damaging the cells, tissues, and/or organs of the subject.

The term "protecting against," as used herein, refers to preventing a subject from developing a bacterial infection (e.g., a Gram-negative bacterial infection) or decreasing the risk that a subject may develop a bacterial infection (e.g., a Gram-negative bacterial infection). Prophylactic drugs used in methods of protecting against a bacterial infection in a subject are often administered to the subject prior to any detection of the bacterial infection. In some embodiments of methods of protecting against a bacterial infection, a subject (e.g., a subject at risk of developing a bacterial infection) may be

administered a compound described herein (e.g., a compound having any one of formulas (l)-(XXXXX)) to prevent the bacterial infection development or decrease the risk of the bacterial infection development.

The term "treating" or "to treat," as used herein, refers to a therapeutic treatment of a bacterial infection (e.g., a Gram-negative bacterial infection) in a subject. In some embodiments, a therapeutic treatment may slow the progression of the bacterial infection, improve the subject's outcome, and/or eliminate the infection. In some embodiments, a therapeutic treatment of a bacterial infection (e.g., a Gram-negative bacterial infection) in a subject may alleviate or ameliorate of one or more symptoms or conditions associated with the bacterial infection, diminish the extent of the bacterial infection, stabilize (i.e., not worsening) the state of the bacterial infection, prevent the spread of the bacterial infection, and/or delay or slow the progress of the bacterial infection, as compare the state and/or the condition of the bacterial infection in the absence of the therapeutic treatment.

The phrase "LPS-induced nitric oxide (NO) production from a macrophage," as used herein, refers to the ability of the lipopolysaccharides (LPS) in Gram-negative bacteria to activate a macrophage and induce NO production from the macrophage. NO production from a macrophage in response to LPS is a signal of macrophage activation, which may lead to sepsis in a subject, e.g., a Gram-negative bacteria infected subject. The disclosure features compounds (e.g., compounds of any one of formulas (l)-(XXXXX)) that are able to bind to LPS in the cell membrane of Gram-negative bacteria to disrupt and permeabilize the cell membrane, thus neutralizing an immune response to LPS (see, e.g., Example 1 1 0). NO production from a macrophage may be measured using available techniques in the art, e.g., a Griess assay, as demonstrated in Example 1 10.

The term "resistant strain of bacteria," as used herein, refers to a strain of bacteria (e.g., Gram- negative or Gram-positive bacteria) that is refractory to treatment with an antibiotic, such as an antibiotic described in Section V of Detailed Description. Antibiotics that a strain of bacteria is resistant to do not include the compounds described herein (e.g., compounds of any one of formulas (l)-(XXXXX)). Resistance may arise through natural resistance in certain types of bacteria, spontaneous random genetic mutations, and/or by inter- or intra-species horizontal transfer of resistance genes. In some embodiments, a resistant strain of bacteria contains a mcr- 1 gene, a mcr-2 gene, and/or a mcr-3 gene. In some embodiments, a resistant strain of bacteria contains a chromosomal mutation conferring polymyxin resistance. In some embodiments, a resistant strain of bacteria contains a mcr- 1 gene, a mcr-2 gene, and/or a mcr-3 gene in combination with other antibiotic resistance genes. In some embodiments, a resistant strain of bacteria is a resistant strain of E. co//^' (e.g., E. coli BAA-2469).

The term "activating an immune cell," as used herein, refers to the ability of a compound to directly or indirectly bind to an immune cell to produce an effective immune response. The ability of a compound to directly or indirectly bind to an immune cell to produce an effective immune response may be quantified by measuring the concentration of the compound at which such immune response is produced. In some embodiments, the concentration of a compound that binds to an immune cell receptor such as dectin-1 or binds to an antibody (e.g., anti-aGal or anti-aRha antibody, which then binds to an immune cell) to trigger an effective immune response may be less than or equal to 10,000 nM as measured in accordance with, e.g., an enzyme-linked immunosorbent assay (ELISA), as in Examples 107 and 108. For instance, an aGal epitope, that binds to an antibody, such as an anti-aGal antibody, may be detected using an ELISA. In an ELISA, a compound containing a particular monosaccharide or oligosaccharide moiety may be immobilized on a support or surface using conventional techniques in the art. After the compound is immobilized to the surface, an antibody that is specific for the particular monosaccharide or

oligosaccharide moiety in the compound is applied over the surface so it is captured by the compound through binding to the monosaccharide or oligosaccharide moiety in the compound. The antibody is often linked to an enzyme (e.g., horseradish peroxidase) for subsequent signal amplification. During signal amplification, the enzyme's substrate (e.g., 3,3'-diaminobenzidine) is added to produce a measurable signal (e.g., color change). In some embodiments, the antibody itself can be detected using a secondary antibody, which is linked to an enzyme.

In some embodiments, the concentration of a compound that binds to an immune cell receptor such as dectin-1 or binds to an antibody (e.g., anti-aGal or anti-aRha antibody, which then binds to an immune cell) to trigger an effective immune response may be less than or equal to 1000 nM or less than or equal to 100 nM as measured in accordance with an ELISA.

The term "innate immune receptor," as used herein, refers to a natural receptor, such as a natural receptor on an immune cell, that binds to a carbohydrate (e.g., a monosaccharide or oligosaccharide moiety) or an optionally substituted carbohydrate and causes a response in the immune system. In some embodiments, an innate immune receptor binds to the monosaccharide or oligosaccharide moiety of the compounds described herein (e.g., compounds of any one of formulas (l)-(XXXXX)). In some embodiments, an innate immune receptor binds to a moiety in Table 2A or 2B.

The term "natural antibody," as used herein, refers to a naturally existing antibody in the circulation of a mammal (e.g., a human) that has not been previously exposed to deliberate immunization. In some embodiments, a natural antibody is an antibody of the immunoglobulin M (IgM) isotype. In some embodiments, a natural antibody binds to the monosaccharide or oligosaccharide moiety of the compounds described herein (e.g., compounds of any one of formulas (l)-(XXXXX)). In some embodiments, a natural antibody binds to a moiety in Table 2A or 2B. In some embodiments, a natural antibody is anti-aGal antibobody or anti-aRha antibody.

The term "subject," as used herein, can be a human, non-human primate, or other mammal, such as but not limited to dog, cat, horse, cow, pig, turkey, goat, fish, monkey, chicken, rat, mouse, and sheep. The term "substantially simultaneously," as used herein, refers to two or more events that occur at the same time or within a narrow time frame of each other. As disclosed herein, a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) and an antibacterial agent (e.g., linezolid or tedizolid) may be administered substantially simultaneously, which means that the compound and the antibacterial agent are administered together (e.g., in one pharmaceutical composition) or separately but within a narrow time frame of each other, e.g., within 10 minutes, e.g., 1 0, 9, 8, 7, 6, 5, 4, 3, 2, or 1 minute, or 45, 30, 15, or 1 0 seconds of each other.

The term "therapeutically effective amount," as used herein, refers to an amount, e.g., pharmaceutical dose, effective in inducing a desired effect in a subject or in treating a subject having a condition or disorder described herein (e.g., a bacterial infection (e.g., a Gram-negative bacterial infection)). It is also to be understood herein that a "therapeutically effective amount" may be interpreted as an amount giving a desired therapeutic and/or preventative effect, taken in one or more doses or in any dosage or route, and/or taken alone or in combination with other therapeutic agents (e.g., an antibacterial agent described herein). For example, in the context of administering a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) that is used for the treatment of a bacterial infection, an effective amount of a compound is, for example, an amount sufficient to prevent, slow down, or reverse the progression of the bacterial infection as compared to the response obtained without administration of the compound.

As used herein, the term "pharmaceutical composition" refers to a medicinal or pharmaceutical formulation that contains at least one active ingredient (e.g., a compound of any one of formulas (I)- (XXXXX)) as well as one or more excipients and diluents to enable the active ingredient suitable for the method of administration. The pharmaceutical composition of the present disclosure includes pharmaceutically acceptable components that are compatible with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)).

As used herein, the term "pharmaceutically acceptable carrier" refers to an excipient or diluent in a pharmaceutical composition. For example, a pharmaceutically acceptable carrier may be a vehicle capable of suspending or dissolving the active compound (e.g., a compound of any one of formulas (I)- (XXXXX)). The pharmaceutically acceptable carrier must be compatible with the other ingredients of the formulation and not deleterious to the recipient. In the present disclosure, the pharmaceutically acceptable carrier must provide adequate pharmaceutical stability to a compound described herein. The nature of the carrier differs with the mode of administration. For example, for oral administration, a solid carrier is preferred; for intravenous administration, an aqueous solution carrier (e.g., WFI, and/or a buffered solution) is generally used.

The term "pharmaceutically acceptable salt," as used herein, represents salts of the compounds described herein (e.g., compounds of any one of formulas (l)-(XXXXX)) that are, within the scope of sound medical judgment, suitable for use in methods described herein without undue toxicity, irritation, and/or allergic response. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in: Pharmaceutical Salts: Properties, Selection, and Use (Eds. P.H. Stahl and C.G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base group with a suitable organic acid.

The term "about," as used herein, indicates a deviation of ±5%. For example, about 1 0% refers to from 9.5% to 10.5%.

Definitions of abbreviations used in the disclosure are provided in Table A below:

Table A

Other features and advantages of the compounds described herein will be apparent from the following detailed description and the claims.

Description of the Drawings

FIG. 1 is a schematic illustrating a 96-well checkerboard synergy MIC plate layout.

FIG. 2 shows the inhibition of nitric oxide (NO) production by Compound 14.

FIGS. 3A-3H show the time-kill analysis for Compound 14 and colistin (COL), respectively, against E. col 7 ATCC 25922, K. pneumoniae ATCC 43816, P. aeruginosa ATCC 27853, and A. baumannii ATCC 17978 at 0, 1 , 2, 4, and 16X CLSI broth microdilution MIC values.

FIG. 4A shows the membrane permeabilization induced by Compound 14, Compound 54a, and

Compound 59, as determined by NPN signal.

FIG. 4B shows the bacterial cell death induced by Compound 14, Compound 54a, and Compound 59, as quantified by SYTOX green signal. FIGS. 5A and 5B show that Rha mediates binding of human Rha antibody to E. coli ATCC 25922-bound Compound 14, 54a, and Compound 59 in an antigen-specific manner. Bound antibodies were detected using either an anti-human IgG secondary antibody (FIG. 5A) or an anti-human IgM secondary antibody (FIG. 5B).

FIG. 5C shows inhibition of Rha-Ab binding to Compound 14 by L-Rha monosaccharide.

FIG. 6 shows Rha-specific binding of purified rabbit Rha antibodies to bacteria.

FIG. 7 shows Rha-specific binding of purified human Rha antibodies to bacteria.

FIGS. 8A-8F show that the killing of E. coli is enhanced by Compound 14, Compound 54a, and

Compound 59 in human blood from donor 1 and donor 2.

FIGS. 9A-9D show that complement dependent cytotoxicity (CDC) is enhanced by Compound 14, Compund 54a, and Compound 59.

FIG. 10 shows that Compound 14 displays rAb-dependent efficacy in a mouse septicemia model.

FIG. 1 1 shows that Compound 14 displays rAb-dependent efficacy in a neutropenic mouse E. coli thigh infection model.

FIG. 12 shows the titration of rAb in the mouse E. coli septicemia model with a fixed dose of

Compound 14.

FIG. 13 shows the metabolic stability of Compound 14 in rat, monkey and human hepatocytes. FIG. 14 shows the titration of Compound 14 in functional potassium channel assays.

FIG. 15 shows the absorbance values at 450 nm for E. coli incubated with Compound 14 and a control compound (Compound 14 without the monosaccharide portion).

Detailed Description

The disclosure features compounds, compositions, and methods for the treatment of bacterial infections (e.g., Gram-negative bacterial infections). The compounds disclosed herein include dimers of cyclic heptapeptides (e.g., two polymyxin cores) linked to each other through a linker and further conjugated to one or more monosaccharide or oligosaccharide moieties. The dimers of cyclic heptapeptides are linked to each other through a linker and/or one or two peptides (e.g., a peptide including a 1 -5 amino acid residue(s)). In particular, the compounds can be used in the treatment of bacterial infections caused by Gram-negative bacteria. In some embodiments, one or more of the compounds described herein are used in combination with an antibacterial agent (e.g., linezolid or tedizolid (e.g., tedizolid phosphate)).

I. Bacterial Infections

Pathogenic bacteria cause bacterial infections and diseases such as tuberculosis, pneumonia, and foodborne illnesses. Bacteria may be categorized into two major types: Gram-positive bacteria and Gram-negative bacteria. Gram-positive bacteria possess a thick cell wall containing multiple layers of peptidoglycan and teichoic acids, while Gram-negative bacteria have a relatively thin cell wall containing fewer layers of peptidoglycan that are surrounded by a second lipid membrane containing

lipopolysaccharides (LPS) and lipoproteins. LPS, also called endotoxins, are composed of

polysaccharides and lipid A. These differences in bacterial ceil wall structure can produce differences in antibiotic susceptibility. Examples of Gram-positive bacteria include, but are not limited to, bacteria in the genus Streptococcus (e.g., Streptococcus pyogenes), bacteria in the genus Staphylococcus (e.g., Staphylococcus cohnii), bacteria in the genus Corynebacterium (e.g., Corynebacterium auris), bacteria in the genus Listeria (e.g., Listeria grayi), bacteria in the genus Bacillus (e.g., Bacillus aerius), and bacteria in the genus Clostridium (e.g., Clostridium acetium). Examples of Gram-negative bacteria include, but are not limited to, bacteria in the genus Escherichia (e.g., Escherichia coli), bacteria in the genus Klebsiella (e.g., Klebsiella granulomatis, Klebsiella oxytoca, Klebsiella pneumoniae, Klebsiella terrigena, and Klebsiella variicola), bacteria in the genus Acinetobacter (e.g., Acinetobacter baumannii,

Acinetobacter calcoaceticus, Acinetobacter kookii, and Acinetobacter junii), bacteria in the genus Pseudomonas (e.g., Pseudomonas aeruginosa), bacteria in the genus Neisseria (e.g., Neisseria gonorrhoeae), bacteria in the genus Yersinia (e.g., Yersinia pestis), bacteria in the genus Vibrio (e.g., Vibrio cholerae), bacteria in the genus Campylobacter (e.g., Campylobacter jejuni), and bacteria in the genus Salmonella (e.g., Salmonella enterica).

Bacteria may evolve to become more or fully resistant to antibiotics. Resistance may arise through natural resistance in certain types of bacteria, spontaneous random genetic mutations, and/or by inter- or intra-species horizontal transfer of resistance genes. Resistant bacteria are increasingly difficult to treat, requiring alternative medications or higher doses, which may be more costly or more toxic. Bacteria resistant to multiple antibiotics are referred to as multidrug resistant (MDR) bacteria. For example, the mcr- 1 gene encodes a phosphoethanolamine transferase (MCR-1 ) which confers resistance to colistin, a natural polymyxin, through modification of LPS. This is the first known horizontally- transferable resistance determinant for the polymyxin class of antibiotics. The mcr- 1 gene has also been found in bacterial strains which already possess resistance to other classes of antibiotics, such as in carbapenem-resistant Enterobacteriaceae (CRE). An mcr- 1 resistance plasmid refers to a bacterial plasmid that carries mcr- 1 alone or in combination with other antibiotic resistance genes. A mcr- 1 resistance plasmid refers to a bacterial plasmid that carries one or more antibiotic resistance genes. Examples of mcr- 1 resistance plasmids include, but are not limited to, pHNSHP45, pMR0516mcr, pESTMCR, pAF48, pAF23, pmcr1 -lncX4, pmcr1 -lncl2, pA31 -12, pVT553, plCBEC72Hmcr, pE15004, pE1 5015, and pE1 5017.

In another example, the mcr-2 gene also confers resistance to colistin. The mcr-2 gene was identified in porcine and bovine colistin-resistance E.coli that did not contain mcr- 1 (Xavier et al., Euro Si/rve/7/ 21 (27), 2016). The mcr-2 gene is a 1 ,617 bp phspoethanolamine transferase harbored on an lncX4 plasmid. The mcr-2 gene has 76.7% nucleotide identity to mcr- 1. Analysis of mcr-2 harboring plasmids from E.coli isolates shows that the mobile element harboring mcr-2 is an IS element of the IS1595 superfamily, which are distinguished by the presence of an ISXC -like transposase domain (Xavier et al., supra). The MCR-2 protein was predicted to have two domains, with domain 1 (1 -229 residues) as a transporter and domain 2 (230-538 residues) as a transferase domain. An mcr-2 resistance plasmid refers to a bacterial plasmid that carries mcr-2 alone or in combination with other antibiotic resistance genes. A mcr-2 resistance plasmid refers to a bacterial plasmid that carries one or more antibiotic resistance genes. Mcr-2 resistance plasmids include, but are not limited to, pKP37-BE and pmcr2-lncX4. In a further example, the mcr-3 gene also confers resistance to colistin. The mcr-3 gene is considered to include variants which encode an MCR-3 protein that confers resistance to cholistin, such as mcr-3 which encodes a protein having an amino acid substitution at V413A. The mcr-3 gene was identified in cholistin-resistant E.coli isolated from swine (Clemente et al., 27^th ECCMID, Vienna, Austria, 2017). The mcr3 gene is harbored on an lncX4 plasmid. Analysis of mcr-3 harboring plasmids from E.coli isolates shows that the mobile element harboring mcr-3 is the IS26 element. A mcr-3 resistance plasmid refers to a bacterial plasmid that carries one or more antibiotic resistance genes. Furthermore, resistant strain E. coli BAA-2469 possesses the New Delhi metallo-p-lactamase (NDM-1 ) enzyme, which makes bacteria resistant to a broad range of β-lactam antibiotics. Additionally, E. coli BAA-2469 is also known to be resistatnt to penicillins (e.g., ticarcillin, ticarcillin/clavulanic acid, piperacillin, ampicillin, and ampicillin/sulbactam), cephalosporins (e.g., cefalotin, cefuroxime, cefuroxime, cefotetan, cefpodoxime, cefotaxime, ceftizoxime, cefazolin, cefoxitin, ceftazidime, ceftriaxone, and cefepime), carbapenems (e.g., doripenem, meropenem, ertapenem, imipenem), quinolones (e.g., nalidixic acid, moxifloxacin, norfloxacin, ciprofloxacin, and levofloxacin), aminoglycosides (e.g., amikacin, gentamicin, and tobramycin), and other antibiotics (e.g., tetracycline, tigecycline, nitrofurantoin, aztreonam,

trimethoprim/sulfamethoxazole).

In some embodiments, a resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene, and/or a chromosomal mutation conferring polymyxin resistance. In some embodiments, a resistant strain of bacteria is a resistant strain of E. coli (e.g., E. coli BAA-2469).

II. Compounds of the Disclosure

Provided herein are synthetic compounds useful in the treatment of bacterial infections (e.g., Gram-negative bacterial infections). The compounds disclosed herein include three components: (i) a first cyclic heptapeptide (e.g., a first polymyxin core), (ii) a second cyclic heptapeptide (e.g., a second polymyxin core), and (iii) one or more monosaccharide or oligosaccharide moieties. The first and second cyclic heptapeptides are linked to each other by way of a linker and/or one or two peptides (e.g., each peptide including a 1 -5 amino acid residue(s)). In some aspects, a cyclic heptapeptide or polymyxin core, as used herein, refers to certain compounds that bind to lipopolysaccharides (LPS) in the cell membrane of Gram-negative bacteria to disrupt and permeabilize the cell membrane, leading to cell death and/or sensitization to other antibiotics. In some aspects, cyclic heptapeptide, as used herein, refers to certain compounds that kill or inhibit the growth of Gram-negative bacteria as determined by measuring the minimum inhibitory concentration (MIC) against at least one Gram-negative bacteria as known in the art, wherein the MIC is 32 μg/mL or less. Cyclic heptapeptides are composed of, at least, amino acid residues, each of which may, independently, have a D- or L- configuration, assembled as a cyclic heptapeptide ring. A cyclic heptapeptide includes seven natural or non-natural amino acid residues attached to each other in a closed ring. The ring contains six bonds formed by linking the carbon in the a- carboxyl group of one amino acid residue to the nitrogen in the a-amino group of the adjacent amino acid residue and one bond formed by linking the carbon in the a-carboxyl group of one amino acid residue to the nitrogen in the γ-amino group in the side chain of the adjacent amino acid residue. For the amino acid residue whose nitrogen in the γ-amino group in the side chain participates in forming the ring, the nitrogen in the a-amino group of this amino acid residue does not participate directly in forming the ring and serves as the linking nitrogen (thus, referred to as the "linking nitrogen" herein) that links one cyclic heptapeptide or polymyxin core to another cyclic heptapeptide or polymyxin core by way of a linker and/or one or two peptides (e.g., a peptide including a 1 -5 amino acid residue(s)). Compounds described herein contain dimers of cyclic heptapeptides (e.g., two polymyxin cores) that are linked to each other at their linking nitrogens through a linker and/or one or two peptides (e.g., a peptide including a 1 -5 amino acid residue(s)), and one or more monosaccharide or oligosaccharide moieties.

In some embodiments, a peptide including one or more (e.g., 1 -5; 1 , 2, 3, 4, or 5) amino acid residues (e.g., natural and/or non-natural amino acid residues) may be covalently attached to the linking nitrogen of the cyclic heptapeptide or the polymyxin core. Cyclic heptapeptides or polymyxin cores may be derived from polymyxins (e.g., naturally existing polymyxins and non-natural polymyxins) and/or octapeptins (e.g., naturally existing octapeptins and non-natural octapeptins). In some embodiments, cyclic heptapeptides may be compounds described in Gallardo-Godoy et al., J. Med. Chem. 59:1068, 2016 (e.g., compounds 1 1 -41 in Table 1 of Gallardo-Godoy et al.), which is incorporated herein by reference in its entirety. Examples of some naturally existing polymyxins and their structures are shown in Table 1 A. Examples of some non-natural polymyxins and their structures are shown in Table 1 B.

Table 1 A. Natural polymyxins and their structures

Table 1 B. Non-natural polymyxins and their structures (Dab: diaminobutyric acid; Dap: diaminopropionic acid; Orn: ornithine;

Abu: 2-aminobutyric acid; NIe: norleucine)

Without being bound by theory, in some aspects, compounds described herein bind to the cell membrane of Gram-negative bacteria (e.g., bind to LPS in the cell membrane of Gram-negative bacteria) to disrupt and permeabilize the cell membrane, leading to cell death and/or sensitization to other antibiotics. In some embodiments, the initial association of the compounds with the bacterial cell membrane occurs through electrostatic interactions between the compounds and the anionic LPS in the outer membrane of Gram-negative bacteria, disrupting the arrangement of the cell membrane.

Specifically, compounds described herein may bind to lipid A in the LPS. More specifically, compounds described herein may bind to one or both phosphate groups in lipid A. In some embodiments, antibiotic- resistant bacteria (e.g., antibiotic-resistant, Gram-negative bacteria) has one phosphate group in lipid A. In some embodiments, compounds described herein may bind to multiple Gram-negative bacterial cells at the same time. The binding of the compounds described herein to the LPS may also displace Mg²⁺ and Ca²⁺ cations that bridge adjacent LPS molecules, causing, e.g., membrane permeabilization, leakage of cellular molecules, inhibition of cellular respiration, and/or cell death. In some embodiments, in addition to disrupting the membrane of bacterial cells, the monosaccharide or oligosaccharide moieties in the compounds serve as a gradient against which immune cells chemotax to the site of bacterial infection and/or growth.

Compounds provided described herein are described by any one of formulas (l)-(XXXXX). In some embodiments, the first and second cyclic heptapeptides are the same. In some embodiments, the first and second cyclic heptapeptides are different. Compounds described herein may be synthesized using available chemical synthesis techniques in the art. In some embodiments, available functional groups in the first and second cyclic heptapeptides, the linker, and the one or more monosaccharide or oligosaccharide moieties, e.g., amines, carboxylic acids, and/or hydroxyl groups, may be used in making the compounds described herein. For example, the linking nitrogen in a cyclic heptapeptide may form an amide bond with the carbon in a carboxylic acid group in the linker. A peptide including one or more (e.g., 1 -3; 1 , 2, or 3) amino acid residues (e.g., natural and/or non-natural amino acid residues) may be also be covalently attached to the linking nitrogen of the cyclic heptapeptide through forming an amide bond between the carbon in a carboxylic acid group in the peptide and the linking nitrogen. In cases where a functional group is not available for conjugation, a molecule may be derivatized using conventional chemical synthesis techniques that are well known in the art. In some embodiments, the compounds described herein contain one or more chiral centers. The compounds include each of the isolated stereoisomeric forms as well as mixtures of stereoisomers in varying degrees of chiral purity, including racemic mixtures. It also encompasses the various diastereomers, enantiomers, and tautomers that can be formed.

The disclosure provides a compound, or a pharmaceutically acceptable salt thereof, described by formula (I):

(E)n

M2 L' M1

(I)

in which wherein M1 includes a first cyclic heptapeptide containing a linking nitrogen and M2 includes a second cyclic heptapeptide containing a linking nitrogen; each E is, independently, a monosaccharide or oligosaccharide moiety; L' is a linker covalently attached to E and to the linking nitrogen in each of M1 and M2; and n is 1 , 2, 3, or 4 (e.g., 1 or 2). In formula (I), each E can, independently, be connected to an atom in L'.

In some embodiments, L' in the compound described by formula (I) is described by formula (L):

(L)

in which L is a remainder of L'; A1 is a 1 -5 amino acid peptide covalently attached to the linking nitrogen in M1 or is absent; and A2 is a 1 -5 amino acid peptide covalently attached to the linking nitrogen in M2 or is absent.

In some embodiments, the disclosure provides a compound, or a pharmaceutically acceptable salt thereof, described by formula (II):

(II)

in which L is a remainder of L'; n is 1 , 2, 3, or 4 (e.g., 1 or 2); each E is, independently, a monosaccharide or oligosaccharide moiety; each of R¹ , R¹², R'¹ , and R'¹² is, independently, a lipophilic moiety, a polar moiety, or H; each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is, independently, optionally substituted C1 -C5 alkamino, a polar moiety, a positively charged moiety, or H; each of R¹⁵ and R'¹⁵ is, independently, a lipophilic moiety or a polar moiety; each of R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R'², R'³, R'⁴, R'⁵, R'⁶, R'⁷, R'⁸, R'⁹, and R'¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; or two R groups on the same or adjacent atoms join to form an optionally substituted 5-8 membered ring; each of a', b', c', a, b, and c is, independently, 0 or 1 ; each of N¹ , N², N³, N⁴, N'¹ , N'², N'³, and N'⁴ is a nitrogen atom; and each of C¹ , C², C³, C'¹ , C'², and C'³ is a carbon atom.

In some embodiments, when R², R³, and C¹ together form an optionally substituted 5-8 membered ring or when R³, R⁴, N¹ , and C¹ together form an optionally substituted 5-8 membered ring, each of R², R³, and R⁴ is, independently, optionally substituted C1 -C20 alkylene or optionally substituted C1 -C20 heteroalkylene.

In some embodiments, when R⁵, R⁶, and C² together form an optionally substituted 5-8 membered ring or when R⁶, R⁷, N², and C² together form an optionally substituted 5-8 membered ring, each of R⁵, R⁶, and R⁷ is, independently, optionally substituted C1 -C20 alkylene or optionally substituted C1 -C20 heteroalkylene.

In some embodiments, when R⁸, R⁹, and C³ together form an optionally substituted 5-8 membered ring or when R⁹, R¹⁰, N³, and C³ together form an optionally substituted 5-8 membered ring, each of R⁸, R⁹, and R¹⁰ is, independently, optionally substituted C1 -C20 alkylene or optionally substituted C1 -C20 heteroalkylene.

In some embodiments, when R'², R'³, and C'¹ together form an optionally substituted 5-8 membered ring or when R'³, R'⁴, N'¹ , and C'¹ together form an optionally substituted 5-8 membered ring, each of R'², R'³, and R'⁴ is, independently, optionally substituted C1 -C20 alkylene or optionally substituted C1 -C20 heteroalkylene.

In some embodiments, when R'⁵, R'⁶, and C'² together form an optionally substituted 5-8 membered ring or when R'⁶, R'⁷, N'², and C'² together form an optionally substituted 5-8 membered ring, each of R'⁵, R'⁶, and R'⁷ is, independently, optionally substituted C1 -C20 alkylene or optionally substituted C1 -C20 heteroalkylene.

In some embodiments, when R'⁸, R'⁹, and C'³ together form an optionally substituted 5-8 membered ring or when R'⁹, R'¹⁰, N'³, and C'³ together form an optionally substituted 5-8 membered ring, each of R'⁸, R'⁹, and R'¹⁰ is, independently, optionally substituted C1 -C20 alkylene or optionally substituted C1 -C20 heteroalkylene.

In some embodiments, the compound of formula (II) has at least one optionally substituted 5-8 membered ring formed by joining (i) R², R³, and C¹ ; (ii) R³, R⁴, N¹ , and C¹ ; (iii) R⁵, R⁶, and C²; (iv) R⁶, R⁷, N², and C²; (v) R⁸, R⁹, and C³; (vi) R⁹, R¹⁰, N³, and C³; (vii) R'², R'³, and C'¹ ; (viii) R'³, R'⁴, N'¹ , and C'¹ ; (ix) R'⁵, R'⁶, and C'²; (x) R'⁶, R'⁷, N'², and C'²; (xi) R'⁸, R'⁹, and C'³; or (xii) R'⁹, R'¹⁰, N'³, and C'³.

In some embodiments, when a is 1 in the compound of formula (II), (i) each of R², R³, and R⁴ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (ii) R², R³, and C¹ together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted cycloalkyl, optionally substituted heterocycloalkyi comprising 1 or 2 heteroatoms independently selected from N, O, and S, and R⁴ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (iii) R³, R⁴, N¹ , and C¹ together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted heterocycloalkyi comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, and R² is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl.

In some embodiments, when b is 1 in the compound of formula (II), (i) each of R⁵, R⁶, and R⁷ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (ii) R⁵, R⁶, and C² together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted cycloalkyl, optionally substituted heterocycloalkyi comprising 1 or 2 heteroatoms independently selected from N, O, and S, and R⁷ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (iii) R⁶, R⁷, N², and C² together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted heterocycloalkyi comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, and R⁵ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

In some embodiments, when c is 1 in the compound of formula (II), (i) each of R⁸, R⁹, and R¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (ii) R⁸, R⁹, and C³ together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted cycloalkyl, optionally substituted heterocycloalkyi comprising 1 or 2 heteroatoms independently selected from N, O, and S, and R¹⁰ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (iii) R⁹, R¹⁰, N³, and C³ together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted heterocycloalkyi comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, and R⁸ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

In some embodiments, when a' is 1 in the compound of formula (II), (i) each of R'², R'³, and R'⁴ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (ii) R'², R'³, and C'¹ together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted cycloalkyl, optionally substituted heterocycloalkyi comprising 1 or 2 heteroatoms independently selected from N, O, and S, and R'⁴ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (iii) R'³, R'⁴, N'¹ , and C'¹ together form a ring (e.g., an optionally substituted 5- 8 membered ring) comprising optionally substituted heterocycloalkyi comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, and R'² is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

In some embodiments, when b' is 1 in the compound of formula (II), (i) each of R'⁵, R'⁶, and R'⁷ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (ii) R'⁵, R'⁶, and C'² together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted cycloalkyl, optionally substituted heterocycloalkyi comprising 1 or 2 heteroatoms independently selected from N, O, and S, and R'⁷ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (iii) R'⁶, R'⁷, N'², and C'² together form a ring (e.g., an optionally substituted 5- 8 membered ring) comprising optionally substituted heterocycloalkyi comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, and R'⁵ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

In some embodiments, when c' is 1 in the compound of formula (II), (i) each of R'⁸, R'⁹, and R'¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (ii) R'⁸, R'⁹, and C'³ together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted cycloalkyl, optionally substituted heterocycloalkyi comprising 1 or 2 heteroatoms independently selected from N, O, and S, and R'¹⁰ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyi, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl; or (iii) R'⁹, R'¹⁰, N'³, and C'³ together form a ring (e.g., an optionally substituted 5-8 membered ring) comprising optionally substituted heterocycloalkyl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, and R'⁸ is a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted alkamino, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted

cycloalkenylene, optionally substituted cycloalkynylene, optionally substituted aryl, optionally substituted heteroalkyl, optionally substituted heterocycloalkyl, optionally substituted heterocycloalkenylene, optionally substituted heterocycloalkynylene, optionally substituted heteroaryl, optionally substituted alkaryl, or optionally substituted heteroalkaryl.

In some embodiments, the disclosure provides a compound, or a pharmaceutically acceptable salt thereof, described by formula (III):

(III)

in which L is a remainder of L'; n is 1 , 2, 3, or 4; each E is, independently, a monosaccharide or oligosaccharide moiety; each of R¹ , R¹², R'¹ , and R'¹² is, independently, a lipophilic moiety; each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is, independently, optionally substituted C1 -C5 alkamino, a polar moiety, or a positively charged moiety; and/or each of R¹⁵ and R'¹⁵ is, independently, a polar moiety.

In some embodiments, a lipophilic moiety is optionally substituted C1 -C20 alkyl, optionally substituted C5-C15 aryl, optionally substituted C6-C35 alkaryl, or optionally substituted C2-C1 5 heteroaryl. In some embodiments, a lipophilic moiety is C1 -C8 alkyl, methyl substituted C2-C4 alkyl, (C1 - C10)alkylene(C6)aryl, phenyl substituted (C1 -C10)alkylene(C6)aryl, or alkyl substituted C4-C9 heteroaryl. In some embodiments, in particular, a lipophilic moiety is benzyl, isobutyl, sec-butyl, isopropyl, n-propyl, methyl, biphenylmethyl, n-octyl, or methyl substituted indolyl.

In some embodiments, optionally substituted C1 -C5 alkamino is CH2CH2NH2.

In some embodiments, a polar moiety includes a hydroxyl group, a carboxylic acid group, an ester group, or an amide group. In some embodiments, a polar moiety is hydroxyl substituted C1 -C4 alkyl. In some embodiments, a polar moiety is CH(CH3)OH.

In some embodiments, a compound provided herein is described by any one of formulas (IV)- (XXXIII). (E)n

In the compounds described herein, the portion L' in a compound of formula (I) and the portion

(E)n

L in a compound of any one of formulas (ll)-(XV) or (XXXIV)-(XXXXX) indicate that one or more (e.g., 1 , 2, 3, or 4; 1 -2, 1 -3, or 1 -4) monosaccharide or oligosaccharide moieties may be attached to L' or L at any atom(s) within L' or L. E represents a monosaccharide or oligosaccharide moiety. The squiggly line

(E)n (E)n in the portion L' and L is not to be construed as a single bond between one or more

monosaccharide or oligosaccharide moieties and an atom in L' or L. In some embodiments, when n is 1 , one monosaccharide or oligosaccharide moiety may be attached to an atom in L' or L (see, e.g., Compounds 1 -4, 7-21 , and 23-25). In some embodiments, when n is 2, two monosaccharide or oligosaccharide moieties may be attached to two atoms in L' or L (see, e.g., Compounds 5, 6, and 22). As described further herein, a linker in a compound described herein (e.g., L' or L) may be a branched structure. As described further herein, a linker in a compound described herein (e.g., L' or L) may be a multivalent structure, e.g., divalent, trivalent, tetravalent, pentavalent, hexavalent, or heptavalent structure, containing two, three, four, five, six, or seven arms, respectively. In some embodiments when the linker has three or more (e.g., three, four, five, six, or seven) arms, two of the arms may be attached to the first and second cyclic heptapeptides and the remaining arm(s) (e.g., remaining one, two, three, four, or five arms) may be attached to one or more (e.g., 1 , 2, 3, 4, or 5; 1 -2, 1 -3, 1 -4, or 1 -5)

(E)n (E)n monosaccaride or oligosaccharide moieties. Thus, L' or L in the portion L' or L may have multiple arms to attached to multiple monosaccharide or oligosaccharide moieties. For example, in Compounds 5, 6, and 22, the linker portion L' or L has four arms, in which two arms are attached to the first and second cyclic heptapeptides and two arms are attached to the first and second monosaccharide moieties (L-Rha).

The disclosure also provides a compound of formula (XVI):

(XVI)

The disclosure also provides a compound of formula (XVII):

(XVII)

The disclosure also provides a compound of formula (XVIII):

(XVIII)

wherein each A¹ and A² is an independently selected amino acid; X is absent or is -CH2CH2C(0)NH-; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; n is 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 10; and e is an integer from 0 to 10; or a

pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XIX):

(XIX)

The disclosure also features a compound of formula (XX):

(XX)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 1 0; e is an integer from 0 to 10; f is an integer from 0 to 10; and g is an integer from 0 to 10; or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXI):

The disclosure also features a compound of formula (XXII):

(XXII)

The disclosure also features a compound of formula (XXIII):

a monosaccharide or an oligosaccharide; Y is

-C(0)CH2CH2-, -CH2-, or is absent; X is -C(0)CH2CH2- or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 15; e is an integer from 1 to 10; f is an integer from 1 to 5; and g is an integer from 1 to 5; or a pharmaceutically acceptable salt thereof. The disclosure also features a compound of formula (XXIV):

(XXIV)

The disclosure also features a compound of formula (XXV):

(XXV)

pharmaceutically acceptable salt thereof. The disclosure also features a compound of formula (XXVI):

(XXVI)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; R is C1 -C20 alkyl; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 20; e is an integer from 0 to 20; and f is an integer from 0 to 20; or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXVII):

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; R is C1 -C20 alkyl; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 20; e is an integer from 1 to 20; and f is an integer from 1 to 20; or a pharmaceutically acceptable salt thereof. The disclosure also features a compound of formula (XXVIII):

(XXVIII)

a monosaccharide or an oligosaccharide; Y is

The disclosure also features a compound of formula (XXIX):

(XXIX)

wherein each A¹ and A² is an independently selected amino acid; each E is independently selected from a monosaccharide or an oligosaccharide; X is -CH2- or -C(O)-; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; and d is an integer from 0 to 15; or a pharmaceutically acceptable salt thereof. The disclosure also features a compound of formula (XXX):

(XXX)

a monosaccharide or an oligosaccharide; Y is

, -C(0)CH₂CH2-, -CH2-, or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 15; g is an integer from 0 to 15; and e and e' are each independently an integer from 1 to 3; or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXI):

(XXXI)

a monosaccharide or an oligosaccharide; each Y is independently selected from

C(0)CH2CH2-, and -CH2-, or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 15; each e is independently from 1 to 15; each f is independently from 1 to 15; and g is from 1 to 15; or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXII):

(XXXII)

a monosaccharide or an oligosaccharide; each Y is independently selected from

C(0)CH₂CH2-, -CH2CH2NHC(0)CH₂CH2-, and -CH2-, or is absent; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 15; each e is independently from 1 to 15; and g is from 1 to 15; or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXIII):

(XXXIII)

, -C(0)CH₂CH₂-, -CH2CH2NHC(0)CH₂CH2-, -C(O)-, and -CH₂-, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 alkenyl, optionally substituted C1 -C20 alkynyl, optionally substituted C1 -C15 aryl, optionally substituted C1 -C15 heteroaryl, or is absent; each Y is independently selected from -CH-, or oxygen; Z is -NH-, -NHC(O)-, oxygen, or sulfur; each m is independently 0, 1 , 2, 3, 4, or 5; each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 15; g is an integer from 1 to 15; e and e' are independently from 1 to 10 and or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXIII), the compound has the formula (XXXII 1- ):

(XXXIII-1 )

or a pharmaceutically acceptable salt thereof.

In some embodiments of a compound of formula (XXXIII), the compound has the formula (XXXIII-

2):

(XXXIII-2)

or a pharmaceutically acceptable salt thereof. The disclosure also features a compound of formula (XXXIV):

(XXXIV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R'², R'³, R'⁴, R'⁵, R'⁶, R'⁷, R'⁸, R'⁹, and R'¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4- C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C1 5 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; or two R groups on the same or adjacent atoms join to form an optionally substituted 5-8 membered ring; each of a', b', c', a, b, and c is, independently, 0 or 1 ; each of N¹ , N², N³, N⁴, N'¹ , N'², N'³, and N'⁴ is a nitrogen atom ; each of C¹ , C², C³, C'¹ , C'², and C'³ is a carbon atom ; L is a linker comprising at least one optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C3-C15 heteroarylene; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXV):

(XXXV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R⁸, R'², R'⁶, and R'⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXV):

(XXXV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R⁸, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 - C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXVI):

(XXXVI)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R⁸, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXVII):

(XXXVII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, and R⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXVIII):

(XXXVIII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXIX):

(XXXIX)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R², R⁶, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXX):

(XXXX) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R² and R⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXXI):

(XXXXI)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each of R² and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXXII):

(XXXXII) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; R² is a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20

heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXXIII):

(XXXXIII)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; each E is, independently, a monosaccharide or oligosaccharide moiety; n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXXIV):

(XXXXIV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; and L comprises an optionally substituted C3 cycloalkylene or an optionally substituted C3 heterocycloalkylene, or a pharmaceutically acceptable salt thereof.

The disclosure also features a compound of formula (XXXXV):

(XXXXV)

The disclosure also features a compound of formula (XXXXVI):

(XXXXVI)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; and L comprises an optionally substituted C5 cycloalkylene or an optionally substituted C5 heterocycloalkylene, or a pharmaceutically acceptable salt thereof.

The disclosure also features a com ound of formula (XXXXVII):

(XXXXVII)

The disclosure also features a com ound of formula (XXXXVIII):

(XXXXVIII)

The disclosure also features a com ound of formula (XXXXIX):

(XXXXIX)

The disclosure also features a compound of formula (XXXXX):

(XXXXX) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl (e.g., CH2CH(CH3)2), or cyclohexylmethyl; and L comprises an optionally substituted C6 arylene, or a pharmaceutically acceptable salt thereof.

ome embodiments of the compounds described herein, E i V 'ΌΗ larmaceutically acceptable salt thereof.

III. Linkers

A linker refers to a linkage or connection between two or more components in a compound (e.g., two cyclic heptapeptides and one or more monosaccharide or oligosaccharide moieties in a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)). In some embodiments, the linker L' in the compound described by formula (I) is described by formula (L):

(L)

in which L is a remainder of L'; A1 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M1 or is absent; and A2 is a 1 -5 amino acid peptide (e.g., a 1 -4, 1 -3, or 1 -2 amino acid peptide) covalently attached to the linking nitrogen in M2 or is absent. In some embodiments, a compound described herein (e.g., compounds of any one of formulas (l)-(XXXXX)) may contain a linker that has a multivalent structure (e.g., divalent, trivalent, tetravalent, pentavalent, hexavalent, or heptavalent structure). In some embodiments, a compound described herein (e.g., compounds of any one of formulas (l)-(XII), (XIV), (XV), and (XXXIII)-(XXXXX); e.g., Compounds 1 -4, 7- 21 , and 23-25) may contain a linker that has a trivalent structure (e.g., a trivalent linker; a linker of formula (L-l); a linker of formual (L-VI)). A trivalent linker has three arms, in which each arm is conjugated to a component of the compound (e.g., a first arm conjugated to a first cyclic heptapeptide, a second arm conjugated to a second heptapeptide, and a third arm conjugated to a monosaccharide or

oligosaccharide moiety). In some embodiments, a compound described herein (e.g., compounds of formula (XIII); Compounds 5, 6, and 22) may contain a linker that has a tetravalent structure (e.g., a tetravalent linker; a linker of formula (L-lla), (L-l lb) , or (L-VII)). A tetravalent linker has four arms, in which each arm is conjugated to a component of the compound (e.g., a first arm conjugated to a first cyclic heptapeptide, a second arm conjugated to a second heptapeptide, a third arm conjugated to a first monosaccharide or oligosaccharide moiety, and a fourth arm conjugated to a second monosaccharide or oligosaccharide moiety). In some embodiments, the one or more monosaccharide or oligosaccharide moieties in the compounds described herein may each be, independently, connected to an atom in the linker. In some embodiments, a compound described herein may contain two or three linkers (e.g., compounds of formula (XIII) or (XIV)), in which each linker has a divalent structure (e.g., a divalent linker; a linker of any one of formulas (L-III)-(L-V)). For example, the first linker may connect the first and second cyclic heptapeptides, the second linker may connect a first monosaccharide or oligosaccharide moiety and a peptide attached to the first cyclic heptapeptide, and a third linker may connect a second monosaccharide or oligosaccharide moiety and a peptide attached to the second cyclic heptapeptide.

In some embodiments, a linker is described by formula (L-l):

L^c

L^B-Q-L^A

(L-l)

wherein L^A is described by formula G^A1-(Z^A1 )_gi-(Y^A1 )hi-(Z^A2)ii-(Y^A2)ji -(Z^A3)ki -(Y^A3)ii -(Z^A4)_mi -(Y^A4)ni-(Z^A5)oi- G^A2; L^B is described by formula G^B1-(Z^B1 )g₂-(Y^B1

L^c is described by

G^A1 is a bond attached to Q in formula (L-l); G^A2 is a bond attached to the first cyclic heptapeptide or a peptide (e.g., a peptide including 1 -5 amino acid residue(s)) attached to the first cyclic heptapeptide, if the peptide is present; G^B1 is a bond attached to Q in formula (L-l); G^B2 is a bond attached to the second cyclic heptapeptide or a peptide (e.g., a peptide including 1 -5 amino acid residue(s)) attached to the second cyclic heptapeptide, if the peptide is present; G^C1 is a bond attached to Q in formula (L-l); G^C2 is a bond attached to at least one monosaccharide or oligosaccharide moiety; each of Z^A1 , Z^A2, Z^A3, Z^A4, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4 and Z^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; each of Y^A1 , Y^A2, Y^A3, Y^A4, Y^B1 , Y^B2, Y^B3, Y^B4, Y^C1 , Y^C2, Y^C3, and Y^C4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C1 5 heteroaryl; each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, and o3 is, independently, 0 or 1 ; Q is a nitrogen atom, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene. Linkers of formula (L-l) that may be used in compounds described herein include, but limited to,

wherein R^* is a bond or comprises one or more of optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, optionally substituted C2-C15 heteroarylene, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, and imino, and

wherein R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl.

In some embodiments, a linker is described by formula (L-lla) or (L-llb):

(L-lla) (L-llb)

wherein L^A is described by formula G^A1-(Z^A1 )_gi-(Y^A1 )hi-(Z^A2)ii-(Y^A2)ji -(Z^A3)ki -(Y^A3)ii -(Z^A4)_mi - (Y^A4)m-(Z^A5)oi-G^A2; L^B is described by formula G^B1-(Z^B1 )_g2-(Y^B1 )h2-(Z^B2)i2-(Y^B2)j2-(Z^B3)_k2-(Y^B3)i2-(Z^B4)m2- (Y^B4)n2-(Z^B5)_o2-G^B2; L^c is described by formula G^c1-(Z^c1 )_g3-(Y^c1 )h3-(Z^C2)i₃-(Y^C2)j3-(Z^C3)k3-(Y^C3)i3-(Z^C4)m3- (Y^C4)n3-(Z^C5)o3-G^C2; L^D is described by formula G^D1 -(Z^D1 )_g4-(Y^D1 )h4-(Z^D2)i₄-(Y^D2)j4-(Z^D3)k4-(Y^D3)i4-(Z^D4)m4- (Y^D4)n4-(Z^D5)o4-G^D2; G^A1 is a bond attached to N^A in formula (L-lla) or N^A in formula (L-llb); G^A2 is a bond attached to the first cyclic heptapeptide or a peptide (e.g., a peptide including 1 -5 amino acid residue(s)) attached to the first cyclic heptapeptide, if the peptide is present; G^B1 is a bond attached to N^A in L of formula (L-lla) or N^A in formula (L-llb); G^B2 is a bond attached to the second cyclic heptapeptide or a peptide (e.g., a peptide including 1 -5 amino acid residue(s)) attached to the second cyclic heptapeptide, if the peptide is present; G^C1 is a bond attached to N^B in formula (L-lla) or C in formula (L-llb); G^C2 is a bond attached to a first monosaccharide or oligosaccharide moiety; G^D1 is a bond attached to N^B in formula (L- lla) or C in formula (L-llb); G^D2 is a bond attached to a second monosaccharide or oligosaccharide moiety; each of Z^A1 Z^A2 Z^A3 Z^A4 Z^A5 Z^B1 Z^B2 Z^B3 Z^B4 Z^B5 Z^C1 Z^C2 Z^C3 Z^C4 Z^C5 Z^D1 Z^D2 Z^D3 Z^D4 and Z^D5 is independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene; each of Y^A1 , Y^A2, Y^A3, Y^A4, Y^B1 , Y^B2, Y^B3, Y^B4, Y^C1 , Y^C2, Y^C3, Y^C4 Y^D1 , Y^D2, Y^D3, and Y^D4 is, independently, O, S, NR\ P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 - C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20

cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; and each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, o3, g4, h4, i4, j4, k4, I4, m4, n4, and o4, is, independently, 0 or 1 . Linkers of formulas (L-lla) and (L-llb) that may be used in compounds described herein include, but are not limited to,

In some embodiments, a linker is described b formula (L-VI):

(L-VI)

wherein L^A is described by formula G^A1-(Z^A1 )_gi-(Y^A1 )hi-(Z^A2)ii-(Y^A2)ji -(Z^A3)ki -(Y^A3)ii -(Z^A4)_mi -(Y^A4)ni-(Z^A5)oi- G^A2; L^B is described by formula G^B1-(Z^B1 )g₂

L^c is described by formula G^c1 -(Z^c1 )_g3-(Y^c1

is a bond attached to Q in formula (L-VI); G^A2 is a bond attached to A1 or M1 if A1 is absent; G^B1 is a bond attached to Q in formula (L-VI); G^B2 is a bond attached to A2 or M2 if A2 is absent; G^C1 is a bond attached to Q in formula (L-VI); G^C2 is a bond attached to E; each of Z^A1 , Z^A2, Z^A3, Z^M, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4, and Z^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2- C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20

heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; each of Y^A1 , Y^A2, Y^A3, Y^A4, Y^B1 , Y^B2, Y^B3, Y^B4, Y^C1 , Y^C2, Y^C3, and Y^C4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C1 5 heteroaryl; each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, and o3 is, independently, 0 or 1 ; Q comprises at least one optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C3-C15 heteroarylene.

In some embodiments, a linker is described b formula (L-VII):

(L-VII)

wherein L^A is described by formula G^A1-(Z^A1 )_gi-(Y^A1 )hi-(Z^A2)ii-(Y^A2)ji -(Z^A3)ki -(Y^A3)ii -(Z^A4)_mi -(Y^A4)ni-(Z^A5)oi - G^A2; L^B is described by formula G^B1-(Z^B1 )_g2-(Y^B1 )h2-(Z^B2)i2-(Y^B2)j2-(Z^B3)_k2-(Y^B3)i2-(Z^B4)m2-(Y^B4)n2-(Z^B5)02-G^B2; L^c is described by formula G^c1 -(Z^c1 )_g3-(Y^c1 )h3-(Z^C2)i₃-(Y^C2)j3-(Z^C3)k3-(Y^C3)i3-(Z^C4)m3-(Y^C4)n3-(Z^C5)03-G^C2^ L^D is described by formula G^D1-(Z^D1 )_g4-(Y^{D 1} )h4-(Z^D2)i₄-(Y^D2)j4-(Z^D3)k4-(Y^D3)i4-(Z^D4)m4-(Y^D4)n4-(Z^D5)o4-G^D2; G^A1 is a bond attached to Q in formula (L-VII); G^A2 is a bond attached to A1 or M1 if A1 is absent; G^B1 is a bond attached to Q in formula (L-VII); G^B2 is a bond attached to A2 or M2 if A2 is absent; G^C1 is a bond attached to Q in formula (L-VII); G^C2 is a bond attached to a first monosaccharide or oligosaccharide moiety, E¹ ; G^D1 is a bond attached to N in formula (L-VII); G^D2 is a bond attached to a second

monosaccharide or oligosaccharide moiety, E²; each of Z^A1 , Z^A2, Z^A3, Z^M, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4, Z^C5, Z^D1 , Z^D2, Z^D3, Z^D4, and Z^D5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene; each of Y^A1 , Y^A2, Y^A3, Y^A4, Y^B1 , Y^B2, Y^B3, Y^B4, Y^C1 , Y^C2, Y^C3, Y^C4, Y^D1 , Y^D2, Y^D3, and Y^D4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2- C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, o3, g4, h4, i4, j4, k4, I4, m4, n4, and o4 is, independently, 0 or 1 ; Q comprises at least one optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C3-C15 heteroarylene.

In some embodiments, a linker provides space, rigidity, and/or flexibility between the two cyclic heptapeptides. In some embodiments, a linker may be a bond, e.g., a covalent bond, e.g., an amide bond, a disulfide bond, a C-N bond, a N-N bond, or any kind of bond created from a chemical reaction, e.g., chemical conjugation. In some embodiments, a linker (Ι_', L, L¹, or L'¹ as shown in any one of formulas (l)-(XVII) and (XXXIV)-(XXXXX)) includes no more than 250 atoms (e.g., 1 -2, 1 -4, 1 -6, 1 -8, 1 -10, 1-12, 1-14, 1-16, 1-18, 1-20, 1-25, 1-30, 1-35, 1-40, 1-45, 1-50, 1-55, 1-60, 1-65, 1-70, 1-75, 1-80, 1-85, 1-90, 1-95, 1-100, 1-110, 1-120, 1-130, 1-140, 1-150, 1-160, 1-170, 1-180, 1-190, 1-200, 1-210, 1-220, 1- 230, 1-240, or 1-250 atom(s); 250, 240, 230, 220, 210, 200, 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 28, 26, 24, 22, 20, 18, 16, 14, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 atom(s)). In some embodiments, a linker (Ι_', L, L¹, or L'¹ as shown in any one of formulas (l)-(XVII) and (XXXIV)-(XXXXX)) includes no more than 250 non-hydrogen atoms (e.g., 1-2, 1-4, 1-6, 1-8, 1-10, 1-12, 1-14, 1-16, 1-18, 1-20, 1-25, 1-30, 1-35, 1-40, 1-45, 1-50, 1-55, 1-60, 1-65, 1-70, 1-75, 1-80, 1-85, 1-90, 1-95, 1-100, 1-110, 1-120, 1-130, 1-140, 1-150, 1-160, 1-170, 1-180, 1-190, 1-200, 1-210, 1- 220, 1 -230, 1 -240, or 1 -250 non-hydrogen atom(s); 250, 240, 230, 220, 210, 200, 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 28, 26, 24, 22, 20, 18, 16, 14, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 non-hydrogen atom(s)). In some embodiments, the backbone of a linker (L\ L, L¹, or L'¹ as shown in any one of formulas (l)-(XVII) and (XXXIV)-(XXXXX)) includes no more than 250 atoms (e.g., 1 -2, 1 -4, 1 -6, 1 -8, 1 -10, 1 -12, 1 -14, 1 -16, 1 -18, 1 -20, 1 -25, 1 -30, 1 -35, 1 -40, 1 - 45, 1-50, 1-55, 1-60, 1-65, 1-70, 1-75, 1-80, 1-85, 1-90, 1-95, 1-100, 1-110, 1-120, 1-130, 1-140, 1-150, 1 -160, 1 -170, 1 -180, 1 -190, 1 -200, 1 -210, 1 -220, 1 -230, 1 -240, or 1 -250 atom(s); 250, 240, 230, 220,

210, 200, 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 95, 90, 85, 80, 75, 70, 65, 60, 55, 50, 45,40, 35, 30, 28, 26, 24, 22, 20, 18, 16, 14, 12, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 atom(s)). The "backbone" of a linker refers to the atoms in the linker that together form the shortest path from one part of the compound to another part of the compound. The atoms in the backbone of the linker are directly involved in linking one part of the compound to another part of the compound. For examples, hydrogen atoms attached to carbons in the backbone of the linker are not considered as directly involved in linking one part of the compound to another part of the compound.

Molecules that may be used to make linkers (Ι_', L, L¹, or L'¹ as shown in any one of formulas (I)- (XVII) and (XXXIV)-(XXXXX)) include at least two functional groups, e.g., two carboxylic acid groups. In some embodiments of a trivalent linker, two arms of a linker may contain two dicarboxylic acids, in which the first carboxylic acid may form a covalent linkage with the first cyclic heptapeptide in the compound and the second carboxylic acid may form a covalent linkage with the second cyclic heptapeptide in the compound, and the third arm of the linker may for a covalent linkage (e.g., a C-0 bond) with one or more monosaccharide or oligosaccharide moieties in the compound. In some embodiments of a divalent linker, the divalent linker may contain two carboxylic acids, in which the first carboxylic acid may form a covalent linkage with one component (e.g., a first cyclic heptapeptide) in the compound and the second carboxylic acid may form a covalent linkage with another component (e.g., a second cyclic heptapeptide) in the compound. In some embodiments, when the first and/or second cyclic heptapeptide is attached to a peptide (e.g., a peptide including a 1 -5 amino acid residue(s)) at the linking nitrogen, the linker may form a covalent linkage with the peptide. In other embodiments of a divalent linker, one end of the linker may form a covalent linkage with the first or second cyclic heptapeptide in the compound and the other end of the linker may form a covalent linkage (e.g., a C-0 bond) with one or more monosaccharide or oligosaccharide moieties.

In some embodiments, dicarboxylic acid molecules may be used as linkers (e.g., a dicarboxylic acid linker). For example, the first carboxylic acid in a dicarboxylic acid molecule may form a covalent linkage with the linking nitrogen of the first cyclic heptapeptide and the second carboxylic acid may form a covalent linkage with the linking nitrogen of the second cyclic heptapeptide. In some embodiments, when the first and/or second cyclic heptapeptide is attached to a peptide (e.g., a peptide including a 1 -5 amino acid residue(s)) at the linking nitrogen, the first carboxylic acid in a dicarboxylic acid linker may form a covalent linkage with the terminal amine group at the end of the peptide that is attached to the first cyclic heptapeptide and the second carboxylic acid in the dicarboxylic acid linker may form a covalent linkage with the terminal amine group at the end of the peptide that is attached to the second cyclic heptapeptide.

mited to,

In some embodiments, dicarboxylic acid molecules, such as the ones described herein, may be further functionalized to contain one or more additional functional groups, which may be used to conjugate to one or more monosaccharide or oligosaccharide moieties.

In some embodiments, a molecule containing one or more sulfonic acid groups may be used to form a linker, in which the sulfonic acid group may form a sulfonamide linkage with the linking nitrogen in a cyclic heptapeptide. In some embodiments, a molecule containing one or more isocyanate groups may be used to form a linker, in which the isocyanate group may form a urea linkage with the linking nitrogen in a cyclic heptapeptide. In some embodiments, a molecule containing one or more haloalkyl groups may be used to form a linker, in which the haloalkyl group may form a covalent linkage, e.g., C-N and C-0 linkages, with a cyclic heptapeptide.

In some embodiments, a linker (Ι_', L, L¹ , or L'¹ as shown in any one of formulas (l)-(XVII) and (XXXIV)-(XXXXX)) may comprise a synthetic group derived from, e.g., a synthetic polymer (e.g., a polyethylene glycol (PEG) polymer). In some embodiments, a linker may comprise one or more amino acid residues. In some embodiments, a linker may be an amino acid sequence (e.g., a 1 -25 amino acid, 1 -10 amino acid, 1 -9 amino acid, 1 -8 amino acid, 1 -7 amino acid, 1 -6 amino acid, 1 -5 amino acid, 1 -4 amino acid, 1 -3 amino acid, 1 -2 amino acid, or 1 amino acid sequence). In some embodiments, a linker (L\ L, L¹ , or L'¹ as shown in any one of formulas (l)-(XVII) and (XXXIV)-(XXXXX)) may include one or more optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene (e.g., a PEG unit), optionally substituted C2-C20 alkenylene (e.g., C2 alkenylene), optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20

In some embodiments, L in a compound of any one of formulas (ll)-(XII) and (XVII) may have a formula of (L-l), (L-lla), or (L-llb). In some embodiments, L in a compound of formula (XIII) may have formula (L-lll), L¹ in a compound of formula (XIII) may have formula (L-IV), and L'¹ in a compound of formula (XIII) may have formula (L-V). In some embodiments, L in a compound of any one of formulas (XXXIV)-(XXXXX) may have a formula of (L-VI) or (L-VII).

Covalent conjugation of two or more components in a compound using a linker may be accomplished using well-known organic chemical synthesis techniques and methods. Complementary functional groups on two components may react with each other to form a covalent bond. Examples of complementary reactive functional groups include, but are not limited to, e.g., amine and activated carboxylic acid, thiol and maleimide, activated sulfonic acid and amine, isocyanate and amine, azide and alkyne, and alkene and tetrazine.

Other examples of functional groups capable of reacting with amino groups include, e.g., alkylating and acylating agents. Representative alkylating agents include: (i) an a-haloacetyl group, e.g., XCH2CO- (where X=Br, CI, or I); (ii) a N-maleimide group, which may react with amino groups either through a Michael type reaction or through acylation by addition to the ring carbonyl group; (iii) an aryl halide, e.g., a nitrohaloaromatic group; (iv) an alkyl halide; (v) an aldehyde or ketone capable of Schiff's base formation with amino groups; (vi) an epoxide, e.g., an epichlorohydrin and a bisoxirane, which may react with amino, sulfhydryl, or phenolic hydroxyl groups; (vii) a chlorine-containing of s-triazine, which is reactive towards nucleophiles such as amino, sufhydryl, and hydroxyl groups; (viii) an aziridine, which is reactive towards nucleophiles such as amino groups by ring opening; (ix) a squaric acid diethyl ester; and (x) an a-haloalkyl ether. Examples of amino-reactive acylating groups include, e.g., (i) an isocyanate and an isothiocyanate; (ii) a sulfonyl chloride; (iii) an acid halide; (iv) an active ester, e.g., a nitrophenylester or N- hydroxysuccinimidyl ester; (v) an acid anhydride, e.g., a mixed, symmetrical, or N-carboxyanhydride; (vi) an acylazide; and (vii) an imidoester. Aldehydes and ketones may be reacted with amines to form Schiff's bases, which may be stabilized through reductive amination.

It will be appreciated that certain functional groups may be converted to other functional groups prior to reaction, for example, to confer additional reactivity or selectivity. Examples of methods useful for this purpose include conversion of amines to carboxyls using reagents such as dicarboxylic anhydrides; conversion of amines to thiols using reagents such as N-acetylhomocysteine thiolactone, S- acetylmercaptosuccinic anhydride, 2-iminothiolane, or thiol-containing succinimidyl derivatives;

conversion of thiols to carboxyls using reagents such as a -haloacetates; conversion of thiols to amines using reagents such as ethylenimine or 2-bromoethylamine; conversion of carboxyls to amines using reagents such as carbodiimides followed by diamines; and conversion of alcohols to thiols using reagents such as tosyl chloride followed by transesterification with thioacetate and hydrolysis to the thiol with sodium acetate.

IV. Monosaccharide or Oligosaccharide Moieties

A monosaccharide moiety is a molecular moiety that has one optionally substituted C6-C9 (exclusive of the substituents) monosaccharide residue. An oligosaccharide moiety is a molecular moiety that includes at least two, e.g., 2-1 50 (e.g., 2-149, 2-140, 2-130, 2-120, 2-1 1 0, 2-100, 2-90, 2-80, 2-70, 2- 60, 2-50, 2-40, 2-30, 2-20, 2-18, 2-16, 2-14, 2-12, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, or 2)) optionally substituted C6-C9 (exclusive of the substituents) monosaccharide residues. In some embodiments, an oligosaccharide moiety includes 2-1 8 optionally substituted C6-C9 (exclusive of the substituents) monosaccharide residues. In some embodiments, an oligosaccharide moiety includes 2-12 optionally substituted C6-C9 (exclusive of the substituents) monosaccharide residues. In some embodiments, an oligosaccharide moiety has a molecule weight that does not exceed 30 kDa, 25 kDa, 24 kDa, 23 kDa, 22 kDa, 21 kDa, 20 kDa, 19 kDa, 1 8 kDa, 17 kDa, 16 kDa, 15 kDa, 14 kDa, 13 kDa, 12 kDa, 1 1 kDa, 10 kDa, 9 kDa, 8 kDa, 7 kDa, 6 kDa, 5 kDa or 3 kDa.

In some embodiments, a monosaccharide moiety has an optionally substituted C6-C9 monosaccharide residue selected from glucose (Glc), galactose (Gal), mannose (Man), allose (All), altrose (alt), gulose (Gul), idose (ido), talose (Tal), fucose (Fuc), rhamnose (Rha; also called L-rhamnose or L-Rha), thia-rhamnose (thia-Rha; also called thia-L-rhamnose or thia-L-Rha), quinovose (Qui), 2- deoxyglucose (2-dGlc), glucosamine (GlcN), galactosamine (GaIN), mannosamine (ManN), fucosamine (FucN), quinovosamine (QuiN), N-Acetyl-glucosamine (GlcNAc); N-Acetyl-galactosamine (GalNAc), N- Acetyl-mannosamine (ManNAc), N-acetyl-fucosamine (FucNAc), N-acetyl-quinovosamine (QuiNAc), glucuronic acid (GlcA), galacturonic acid (GalA), mannuronic acid (ManA), iduronic acid (IdoA), sialic acid (Sia), neuraminic acid (Neu), N-Acetyl-neuraminic acid (Neu5Ac), N-Glycolyl-neuraminic acid (Neu5Gc), glucitol (Glc-ol), galactitol (Gal-ol), mannitol (Man-ol), fructose (Fru), sorbose (Sor), tagatose (Tag), thevetose (The), acofriose (Aco), digitoxose (Dig), cymarose (Cym), abequose (Abe), colitose (Col), tyvelose (Tyv), ascarylose (Asc), paratose (Par), or N-acetyl-muramic acid (MurNAc). In some embodiments, a monosaccharide moiety has an optionally substituted C6-C9 monosaccharide residue selected from Rha, Gal, Glc, GIcA (Glucuronic acid), GlcNAc, GalNAc, GlcN(Gc) (N-Glycolyl-Glucosamine), Neu5Ac, Neu5Gc (N-Glycolyl-neuraminic acid), Fuc, Man, -htePCbMan (mannose phosphate), 6-H2PO3GIC (glucose phosphate), Mur (muramoyl), Mur-L-Ala-D-i-Gln-Lys, (Mur)- 3-O-GlcNAc, sulfate-galactose (Su-Gal), disulfate-galactose (Su2-Gal), sulfate-glucose (Su-Glc), sulfate- GlcNAc (Su-GlcNAc), or sulfate-GalNAc (Su-GalNAc).

Rhamnose (Rha) occurs in nature in its L-form, thus, rhamnose is also referred to as L-rhamnose or L-Rha. Rhamnose may be in a form (also called a-Rha or a-L-Rha) or β form (also called β-Rha or β- L-Rha). Thia-rhamnose has -SH attached at the anomeric carbon instead of -OH. Thia-rhamnose is also referred to as thia-Rha or thia-L-Rha. Thia-rhamnose may be in a form (also called thia-a-Rha or thia-a-L-Rha) or in β form (also called thia^-Rha or thia^-L-Rha). The structures of Rha, a-Rha, and β-Rha are shown below. In some embodiments, the monosaccharide moiety is an optionally substituted a-Rha or an optionally substituted thia-a-Rha. In some embodiments, a compounds described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) may contain a-Rha.

In some embodiments, in an oligosaccharide moiety, each of the optionally substituted C6-C9 monosaccharide residues may, independently, joined to an adjacent monosaccharide residue through an O-glycosidic, S-glycosidic, N-glycosidic linkage, or C-glycosidic. The binding at an O-glycosidic, S- glycosidic, N-glycosidic, or C-glycosidic linkage may be an a- or β-configuration, for example, through 1 ,2- , 1 ,3-, 1 ,4-, 1 ,6-, 2,3-, 2,6-, or 2,8-linkage, or a linkage such as 3-0, for example, a1 -2, a1 -3, a1 -4, a1 -6, a2-3, a2-6, a2-8, β1 -2, β1 -3, β1 -4, or β1 -6. In some embodiments, each of the optionally substituted C6- C9 monosaccharide residues is, independently, glucose (Glc), galactose (Gal), mannose (Man), allose (All), altrose (alt), gulose (Gul), idose (ido), talose (Tal), fucose (Fuc), rhamnose (Rha), quinovose (Qui), 2-deoxyglucose (2-dGlc), glucosamine (GlcN), galactosamine (GaIN), mannosamine (ManN), fucosamine (FucN), quinovosamine (QuiN), N-Acetyl-glucosamine (GlcNAc); N-Acetyl-galactosamine (GalNAc), N- Acetyl-mannosamine (ManNAc), N-acetyl-fucosamine (FucNAc), N-acetyl-quinovosamine (QuiNAc), glucuronic acid (GIcA), galacturonic acid (GalA), mannuronic acid (ManA), iduronic acid (IdoA), sialic acid (Sia), neuraminic acid (Neu), N-Acetyl-neuraminic acid (Neu5Ac), N-Glycolyl-neuraminic acid (Neu5Gc), glucitol (Glc-ol), galactitol (Gal-ol), mannitol (Man-ol), fructose (Fru), sorbose (Sor), tagatose (Tag), thevetose (The), acofriose (Aco), digitoxose (Dig), cymarose (Cym), abequose (Abe), colitose (Col), tyvelose (Tyv), ascarylose (Asc), paratose (Par), or N-acetyl-muramic acid (MurNAc). In some embodiments, each of the optionally substituted C6-C9 monosaccharide residues is, independently, Rha, Gal, Glc, GIcA (Glucuronic acid), GlcNAc, GalNAc, GlcN(Gc) (N-Glycolyl-Glucosamine), Neu5Ac, Neu5Gc (N-Glycolyl-neuraminic acid), Fuc, Man, -H2P03Man (mannose phosphate), 6-H2PO3GIC

(glucose phosphate), Mur (muramoyl), Mur-L-Ala-D-i-Gln-Lys, (Mur)-3-0-GlcNAc, sulfate-galactose (Su- Gal), disulfate-galactose (Su2-Gal), sulfate-glucose (Su-Glc), sulfate-GlcNAc (Su-GlcNAc), or sulfate- GalNAc (Su-GalNAc). In some embodiments, an oligosaccharide moiety may be straight or branched. In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety is substituted with one or more, such as 1 -3, substituents independently selected from sulfate, phosphate, methyl, acetyl, naturally amino acids, and non-naturally occurring amino acids.

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety is substituted with one or more, such as 1 -3, substituents independently selected from sulfate, phosphate, methyl, acetyl.

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety is substituted with one or more, such as 1 -3, substituents independently selected from naturally occurring amino acids and non- naturally occurring amino acids. If the monosaccharide residue is linked to an amino acid, such as serine, the linkage may be, for example, a a1 -O linkage. If the linkage is through a sulfate, the linkage may be through a hydroxyl group, for example, a 2-0, 3-0 , 4-0, or 6-0 linkage, such as 3-0-Su-Gal, (6- 0-Su)Gal, (6-0-Su)Glc, 6-0-Su-GlcNAc, (6-0-Su)GalNAc, 3,6-0-Su2-Gal, 3,4-0-Su2-Gal, or 4-0-Su-Gal. Naturally occurring amino acids include Ala, Arg, Asn, Asp, Cys, Gin, Glu, Gly, His, He, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, and Val. A non-naturally occurring amino acid" is an amino acid that is not naturally produced or found in a mammal. Examples of non-naturally occurring amino acids include D- amino acids; an amino acid having an acetylaminomethyl group attached to a sulfur atom of a cysteine; a pegylated amino acid; the omega amino acids of the formula NH2(CH2)nCOOH where n is 2-6, neutral nonpolar amino acids, such as sarcosine, t-butyl alanine, t-butyl glycine, N-methyl isoleucine, and norleucine; oxymethionine; phenylglycine; citrulline; methionine sulfoxide; cysteic acid; ornithine;

diaminobutyric acid; and hydroxyproline. Other amino acids are a-aminobutyric acid, a-amino-a- methylbutyrate, aminocyclopropane-carboxylate, aminoisobutyric acid, aminonorbornyl-carboxylate, L- cyclohexylalanine, cyclopentylalanine, L-N-methylleucine, L-N-methylmethionine, L-N-methylnorvaline, L- N-methylphenylalanine, L-N-methylproline, L-N-methylserine, L-N-methyltryptophan, D-ornithine, L-N- methylethylglycine, L-norleucine, a-methyl-aminoisobutyrate, a-methylcyclohexylalanine, D-a- methylalanine, D-a-methylarginine, D-a-methylasparagine, D-a-methylaspartate, D-a-methylcysteine, D- a-methylglutamine, D-a-methylhistidine, D-a-methylisoleucine, D-a-methylleucine, D-a-methyllysine, D-a- methylmethionine, D-a-methylornithine, D-a-methylphenylalanine, D-a-methylproline, D-a-methylserine, D-N-methylserine, D-a-methylthreonine, D-a-methyltryptophan, D-a-methyltyrosine, D-a-methylvaline, D- N-methylalanine, D-N-methylarginine, D-N-methylasparagine, D-N-methylaspartate, D-N-methylcysteine, D-N-methylglutamine, D-N-methylglutamate, D-N-methylhistidine, D-N-methylisoleucine, D-N- methylleucine, D-N-methyllysine, N-methylcyclohexylalanine, D-N-methylornithine, N-methylglycine, N- methylaminoisobutyrate, N-(1 -methylpropyl)glycine, N-(2-methylpropyl)glycine, D-N-methyltryptophan, D- N-methyltyrosine, D-N-methylvaline, γ-aminobutyric acid, L-t-butylglycine, L-ethylglycine, L- homophenylalanine, L-a-methylarginine, L-a-methylaspartate, L-a-methylcysteine, L-a-methylglutamine, L-a-methylhistidine, L-a-methylisoleucine, L-a-methylleucine, L-a-methylmethionine, L-a-methylnorvaline, L-a-methylphenylalanine, L-a-methylserine, L-a-methyltryptophan, L-a-methylvaline, N-(N-(2,2- diphenylethyl) carbamylmethylglycine, 1 -carboxy-1 -(2,2-diphenyl-ethylamino) cyclopropane, 4- hydroxyproline, ornithine, 2-aminobenzoyl (anthraniloyl), D-cyclohexylalanine, 4-phenyl-phenylalanine, L- citrulline, a-cyclohexylglycine, L-1 ,2,3,4-tetrahydroisoquinoline-3-carboxylic acid, L-thiazolidine-4- carboxylic acid, L-homotyrosine, L-2-furylalanine, L-histidine (3-methyl), N-(3-guanidinopropyl)glycine, O- methyl-L-tyrosine, O-glycan-serine, meta-tyrosine, nor-tyrosine, L-N,N',N"-trimethyllysine, homolysine, norlysine, N-glycan asparagine, 7-hydroxy-1 ,2,3,4-tetrahydro-4-fluorophenylalanine, 4- methylphenylalanine, bis-(2-picolyl)amine, pentafluorophenylalanine, indoline-2-carboxylic acid, 2- aminobenzoic acid, 3-amino-2-naphthoic acid, asymmetric dimethylarginine, L-tetrahydroisoquinoline-1 - carboxylic acid, D-tetrahydroisoquinoline-1 -carboxylic acid, 1 -amino-cyclohexane acetic acid, D/L- allylglycine, 4-aminobenzoic acid, 1 -amino-cyclobutane carboxylic acid, 2 or 3 or 4-aminocyclohexane carboxylic acid, 1 -amino-1 -cyclopentane carboxylic acid, 1 -aminoindane-1 -carboxylic acid, 4-amino- pyrrolidine-2-carboxylic acid, 2-aminotetraline-2-carboxylic acid, azetidine-3-carboxylic acid, 4-benzyl- pyrrolidine-2-carboxylic acid, tert-butylglycine, b-(benzothiazolyl-2-yl)-alanine, b-cyclopropyl alanine, 5,5- dimethyl-1 ,3-thiazolidine-4-carboxylic acid, (2R,4S)4-hydroxypiperidine-2-carboxylic acid, (2S,4S) and (2S,4R)-4-(2-naphthylmethoxy)-pyrrolidine-2-carboxylic acid, (2S,4S) and (2S,4R)4-phenoxy-pyrrolidine- 2-carboxylic acid, (2R,5S)and(2S,5R)-5-phenyl-pyrrolidine-2-carboxylic acid, (2S,4S)-4-amino-1 -benzoyl- pyrrolidine-2-carboxylic acid, t-butylalanine, (2S,5R)-5-phenyl-pyrrolidine-2-carboxylic acid, 1 - aminomethyl-cyclohexane-acetic acid, 3,5-bis-(2-amino)ethoxy-benzoic acid, 3,5-diamino-benzoic acid, 2- methylamino-benzoic acid, N-methylanthranylic acid, L-N-methylalanine, L-N-methylarginine, L-N- methylasparagine, L-N-methylaspartic acid, L-N-methylcysteine, L-N-methylglutamine, L-N- methylglutamic acid, L-N-methylhistidine, L-N-methylisoleucine, L-N-methyllysine, L-N-methylnorleucine, L-N-methylornithine, L-N-methylthreonine, L-N-methyltyrosine, L-N-methylvaline, L-N-methyl-t- butylglycine, L-norvaline, a-methyl-Y-aminobutyrate, 4,4'-biphenylalanine, a-methylcylcopentylalanine, a- methyl-a-napthylalanine, a-methylpenicillamine, N-(4-aminobutyl)glycine, N-(2-aminoethyl)glycine, N-(3- aminopropyl)glycine, N-amino-a-methylbutyrate, a-napthylalanine, N-benzylglycine, N-(2- carbamylethyl)glycine, N-(carbamylmethyl)glycine, N-(2-carboxyethyl)glycine, N-(carboxymethyl)glycine, N-cyclobutylglycine, N-cyclodecylglycine, N-cycloheptylglycine, N-cyclohexylglycine, N-cyclodecylglycine, N-cylcododecylglycine, N-cyclooctylglycine, N-cyclopropylglycine, N-cycloundecylglycine, N-(2,2- diphenylethyl)glycine, N-(3,3-diphenylpropyl)glycine, N-(3-guanidinopropyl)glycine, N-(1 - hydroxyethyl)glycine, N-(hydroxyethyl))glycine, N-(imidazolylethyl))glycine, N-(3-indolylyethyl)glycine, N- methyl-Y-aminobutyrate, D-N-methylmethionine, N-methylcyclopentylalanine, D-N-methylphenylalanine, D-N-methylproline, D-N-methylthreonine, N-(1 -methylethyl)glycine, N-methyl-napthylalanine, N- methylpenicillamine, N-(p-hydroxyphenyl)glycine, N-(thiomethyl)glycine, penicillamine, L-a-methylalanine, L-a-methylasparagine, L-a-methyl-t-butylglycine, L-methylethylglycine, L-a-methylglutamate, L-a- methylhomophenylalanine, N-(2-methylthioethyl)glycine, L-a-methyllysine, L-a-methylnorleucine, L-a- methylornithine, L-a-methylproline, L-a-methylthreonine, L-a-methyltyrosine, L-N-methyl- homophenylalanine, N-(N-(3,3-diphenylpropyl) carbamylmethylglycine, L-pyroglutamic acid, D- pyroglutamic acid, O-methyl-L-serine, O-methyl-L-homoserine, 5-hydroxylysine, a-carboxyglutamate, phenylglycine, L-pipecolic acid (homoproline), L-homoleucine, L-lysine (dimethyl), L-2-naphthylalanine, L- dimethyldopa or L-dimethoxy-phenylalanine, L-3-pyridylalanine, L-histidine (benzoyloxymethyl), N- cycloheptylglycine, L-diphenylalanine, O-methyl-L-homotyrosine, L-p-homolysine, O-glycan-threoine, Ortho-tyrosine, L-N.N'-dimethyllysine, L-homoarginine, neotryptophan, 3-benzothienylalanine, isoquinoline-3-carboxylic acid, diaminopropionic acid, homocysteine, 3,4-dimethoxyphenylalanine, 4- chlorophenylalanine, L-1 ,2,3,4-tetrahydronorharman-3-carboxylic acid, adamantylalanine, symmetrical dimethylarginine, 3-carboxythiomorpholine, D-1 ,2,3,4-tetrahydronorharman-3-carboxylic acid, 3- aminobenzoic acid, 3-amino-1 -carboxymethyl-pyridin-2-one, 1 -amino-1 -cyclohexane carboxylic acid, 2- aminocyclopentane carboxylic acid, 1 -amino-1 -cyclopropane carboxylic acid, 2-aminoindane-2-carboxylic acid, 4-amino-tetrahydrothiopyran-4-carboxylic acid, azetidine-2-carboxylic acid, b-(benzothiazol-2-yl)- alanine, neopentylglycine, 2-carboxymethyl piperidine, b-cyclobutyl alanine, allylglycine, diaminopropionic acid, homo-cyclohexyl alanine, (2S,4R)- 4-hydroxypiperidine-2-carboxylic acid, octahydroindole-2- carboxylic acid, (2S,4R) and (2S,4R)-4-(2-naphthyl), pyrrolidine-2-carboxylic acid, nipecotic acid, (2S,4R)and (2S,4S)-4-(4-phenylbenzyl) pyrrolidine-2-carboxylic acid, (3S)-1 -pyrrolidine-3-carboxylic acid, (2S,4S)-4-tritylmercapto-pyrrolidine-2-carboxylic acid, (2S,4S)-4-mercaptoproline, t-butylglycine, N,N- bis(3-aminopropyl)glycine, 1 -amino-cyclohexane-1 -carboxylic acid, N-mercaptoethylglycine, and selenocysteine. In some embodiments, amino acid residues may be charged or polar. Charged amino acids include alanine, lysine, aspartic acid, or glutamic acid, or non-naturally occurring analogs thereof. Polar amino acids include glutamine, asparagine, histidine, serine, threonine, tyrosine, methionine, or tryptophan, or non-naturally occurring analogs thereof.

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may be in a closed ring form

(i.e., where the aldehyde/ketone carbonyl carbon (C=0) and hydroxyl group (-OH) in the open-chain monosaccharide residue reacts to form a hemiacetal with a new C-O-C bridge) or an open ring form.

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety is an optionally substituted C6 monosaccharide residue.

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety is an optionally substituted C9 monosaccharide residue, e.g., sialic acid (Sia), Neuraminic acid (Neu), N-Acetyl-neuraminic acid

(Neu5Ac), or N-Glycolyl-neuraminic acid (Neu5Gc).

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may include a hexose residue, e.g., glucose (Glc), galactose (Gal), mannose (Man), allose (All), altrose (alt), gulose (Gul), idose

(ido), or talose (Tal).

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may include a deoxyhexose residue, e.g., a hexose residue without the hydroxyl group at the 6-position or the 2-position, e.g., fucose (Fuc), rhamnose (Rha), quinovose (Qui), or 2-deoxyglucose (2-dGlc).

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may also include an aminohexose residue, e.g., a hexose residue with an amino group or an N-acetylated amino group at the 2 -position, e.g., glucosamine (GlcN), galactosamine (GaIN), mannosamine (ManN), fucosamine (FucN), quinovosamine (QuiN), N-Acetyl-glucosamine (GlcNAc), N-Acetyl-galactosamine (GalNAc), N-Acetyl- mannosamine (ManNAc), N-acetyl-fucosamine (FucNAc), or N-acetyl-quinovosamine (QuiNAc).

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may include a uronic acid residue, e.g., a hexose residue with a negatively charged carboxylate at the 6-position, e.g., glucuronic acid (GlcA), galacturonic acid (GalA), mannuronic acid (ManA), or iduronic acid (IdoA).

In addition to hexose residues, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may include a sialic acid residue, e.g., a residue of a nine-carbon acidic sugar, e.g., a residue of. Sialic acid (Sia), Neuraminic acid (Neu), N-Acetyl-neuraminic acid (Neu5Ac), or N-Glycolyl-neuraminic acid (Neu5Gc).

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may include a sugar alcohol, e.g., glucitol (Glc-ol), galactitol (Gal-ol), or mannitol (Man-ol).

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may include other compounds, e.g., thevetose (The), acofriose (Aco), digitoxose (Dig), cymarose (Cym), abequose (Abe), colitose (Col), tyvelose (Tyv), ascarylose (Asc), paratose (Par), and N-acetyl-muramic acid (MurNAc).

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety is Gal, such as aGal (for example, a1 -3Gal). An aGal epitope, i.e., an oligosaccharide moiety that exhibits specific binding to an anti-aGal antibody, may also include a moiety comprising an a-D-galactopyranoside moiety, Gal, Galal-3Gal, Galal-4Gal, Galal-6Gal, Galal-3Gala1 -3GlcNAc, Galal-3Gala1 -4Gal, Galal-3Gala1 -4GlcNAc, Galal-3Gala1 -4Glc, Galal-3Gala1 -4[3-deoxy]GlcNAc, Galal-3Gala1 -4[6-deoxy]GlcNAc, Galal-3Gala1 - 4Gala1 -3Gal, Galal-3Gala1 -4GlcNAca1 -3Gala1 -4Glc, and any multimers and combinations thereof. Galal-3Gal 1 -4GlcNAc-R (aGal epitope) glyconjugates have been reported in Macher, et al. Biochim Biophys Acta 1780: 75-88, 2008.

Exemplary aGal epitopes that may be included in a compound described here (e.g., a compound of any one of formulas (l)-(XXXXX)) are shown below:

If present, optional substituents may include 1 -3 substituents independently selected from sulfate, phosphate, methyl, acetyl, naturally occurring amino acid residues, and non-naturally occurring amino acid residues on each monosaccharide residue. These may be linked through an O, S, or N, such as a sulfamate linkage through an S. The amino acid residues may be charged or polar and includes isomers thereof. Charged amino acids include alanine, lysine, aspartic acid, or glutamic acid. Polar amino acids include glutamine, asparagine, histidine, serine, threonine, tyrosine, methionine or tryptophan. For example, the amino acid substituent, if present, may be alanine, lysine, serine, glutamine, or i-glutamine, or asparagine.

In some embodiments, the optionally substituted monosaccharide residue(s) (i.e., at least one monosaccharide residue) in the monosaccharide or oligosaccharide moiety may contain at least one (such as 1 -12) of the following optionally substituted monosaccharide residues: Rha, Gal, Glc, GlcA (Glucuronic acid), GlcNAc, GalNAc, GlcN(Gc) (N-Glycolyl-Glucosamine), Neu5Ac, Neu5Gc (N-Glycolyl- neuraminic acid), Fuc, Man, -htePCbMan (mannose phosphate), 6-H2PO3GIC (glucose phosphate), Mur (muramoyl), Mur-L-Ala-D-i-Gln-Lys, (Mur)-3-0-GlcNAc, sulfate-galactose (Su-Gal), disulfate-galactose (Su2-Gal), sulfate-glucose (Su-Glc), sulfate-GlcNAc (Su-GlcNAc), or sulfate-GalNAc (Su-GalNAc).

In some embodiments, the 1 -12 optionally substituted C6-C9 monosaccharide residues in the monosaccharide or oligosaccharide moiety may be β-glucan residues. In some embodiments, the β- Glucan residue includes 2-12 optionally substituted glucopyranosyl monosaccharide residues each independently joined to an adjacent glucopyranosyl monosaccharide residue via an O-glycosidic, S- glycosidic, N-glycosidic, or C-glycosidic, linkage to form β-linked chains, which retain the ability to bind dectin-1 . Optional substituents may include, e.g., 1 -3 substituents, such as with acetyl, sulfate, phosphate, or a natural or non-natural amino acid. Some examples of β-glucan residues include thia-β- glucan residues such as the structures shown below:

In some embodiments, an oligosaccharide moiety may be a β-glucan having more than 10 monosaccharides and/or a molecular weight of at least 3 kDa (e.g., 3-30 kDa; e.g., 3-29, 3-25, 3-20, 3-15, 3-10, 3-8, 3-6, 3-5, or 3-4 kDa). For example, an oligosaccharide moiety may be a β(1→3, 1→6)-glucan, e.g., laminarin, or a β(1→6)-glucan, e.g., pustulan.

In some embodiments, a monosaccharide or oligosaccharide moiety directly or indirectly activates an immune cell. In some embodiments, the monosaccharide or oligosaccharide moiety directly binds an immune cell. For example, β-glucans bind dectin-1 receptors. When bound to dectin-1 , which internalizes the β-glucan, β-glucan mediates the production of reactive oxygen species (ROS), activation of NF- B, and subsequent secretion of proinflammatory cytokines. The β-giucan receptor, dectin-1 , is predominantly expressed on the surface of cells of monocytes/macrophages and neutrophils. In some embodiments, the monosaccharide or oligosaccharide moiety indirectly binds an immune cell. For example, without being bound by theory, the monosaccharide or oligosaccharide moiety may bind to an antibody. The antibody in turn may bind to Fc receptors found on the surface of certain on immune cells including, among others, B lymphocytes, follicular dendritic cells, natural killer cells, macrophages, neutrophils, eosinophils, basophils, and mast cells. For example, aGal or aRha will bind to anti-aGal or anti-aRha antibody, respectively, thereby indirectly activating immune cells.

In some embodiments, a monosaccharide or oligosaccharide moiety is a ligand to an innate immune receptor. In some embodiments, the innate immune receptor is AICL, BDCA2, CLEC2, Complement receptor 3, Complement receptor 4, DCIR, dectin-1 , dectin-2, DC-SIGN, a C-Type lectin receptor, MMR, langerin, TLR2, Mincle, MBL, or KCR. In some embodiments, the monosaccharide or oligosaccharide moiety binds to an antibody. In some embodiments, the antibody is a natural antibody. In some embodiments, the natural antibody is an antibody of the immunoglobulin M (IgM) isotype.

Glycans bound by antibodies contained in intravenous immunoglobulin (IVIG) are studied by Gunten et al., J. Allergy Clin Immunol. 123:1268, 2009, e.g., see Tables I and E1 of Gunten et al., which is incorporated herein by reference in its entirety. In some embodiments, a monosaccharide or oligosaccharide moiety in a compound described herein (e.g., a compound of any one of formulas (I)- (XXXXX)) may be any one of the glycans listed in Tables I and E1 of Gunten et al. Examples of glycans studied by Gunten et al. are shown in Table 2A. Anti-carbohydrate antibodies found in normal sera have been studied by Huflejt et al., Molecular Immunology 46:3037-3049, 2009, using a library of glycans shown in Table 2B.

Table 2A

Structure of Glycan Structure of Glycan

Neu5Aca2-3Galp1 -3GlcNAcp1 -3Galp1 -

1 GlcNAcpl -2Gcrtp1 -3GalNAca 10

3GlcNAcp

NeuAca2-6Galp1 -4GlcNAcp1 -3Galp1 -

2 GlcNAcal -3Galp1 -4GlcNAcp 1 1

3GlcNAcp

GlcNAcpl -3(GlcNAcp1 -4)(GlcNAcp1 -

3 12 a-L-Rha

6)GlcNAc

4 GlcNAcpl -4MDPLys 13 [su]3Galp1 -3GlcNAcp

5 GlcNAcpl -4(GlcNAcp1 -6)GalNAca 14 HOOC(CH3)CH-3-0-GlcNAcp1 -4GlcNAcp

6 GlcNAcpl -4Galp1 -4GlcNAcp Galpl -3(Fuca1 -4)GlcNAcp1 -2Mana1 - 3[Galp

7 Gala1 -3Galp1 -3GlcNAcp 15

1 -3(Fuca1 -4)GlcNAcp1 -2Mana1 -6]Man p1 -4GlcNAcp1 -4GlcNAcp

8 NeuAc(9Ac)a2-3Galp1 -3GlcNAcp

Neu5Aca2-3Galp1 -4GlcNAcp1 -3Galp1 -

9

3GlcNAcp

Table 2B

(Gc: glyc ;olyl; GIcA: glucuronic acid; Mur: muramoyi; O S: oligo saccharide; P: phosphate; Ser. Serine; Sia: Neu5Ac Su. Sulfate.)

oiruciure oi uiycan oiruciure οτ iycan

1 Neu5Aca2-6Galp1 -4GlcNAcp- 80 GlcNAcpl -3Galp1 -3GalNAca- Structure of Glycan Structure of Glycan

Galp1-4GlcNAcp1-3

Galp1-4GlcNAcp- 81 GalNAcal -3(Fuca1 -2)Galp1 -4GlcNAcp- Galp1-4GlcNAcp1-6

Neu5Gca2-6Galp1 -4GlcNAcp- 82 GalNAca1-0-Ser

Fuca1-3

GlcNAcp- 83 GlcNAcpl -6Galp1 -4GlcNAcp- Neu5Aca2-3Galp1-4

Mana1-3

Manal -6(Mana1 -3)Manp- 84 GlcNAcpl -6(Galp1 -3)GalNAca- Mana1-6

Manal -3

Manpl -4GlcNAcp1 -4GlcNAcp- 85 GlcNAcp1-3Galp1-4Glcp- Mana1-6

Neu5Aca2-8Neu5Aca2- 86 Galpl -4GlcNAcp1 -3GalNAca-

Galpl -4(Fuca1 -3)GlcNAcp- 87 GlcAp1-3Galp-

3-0-Su-Galp1 -4(Fuca1 -3)GlcNAcp- 88 Neu5Aca2-8Neu5Acp-OCH2C₆H₄-p-

Fuca1-2Galp1-4GlcNAcp- 89 Neu5Aca2-3Galp1 -4Glcp-

Fuca1-3

6-O-Su-GlcNAcp- 90 Fuca1-2Galp- Neu5Aca2-3Galp1-4

Neu5Aca2-3Galp1 -3(Fuca1 -4)GlcNAcp- 91 3,4-0-Su₂-Galp1 -4GlcNAcp-

GlcNAcp1-6

Galp1-4GlcNAc- 92 Neu5Aca2-6Galp- Galp1-4GlcNAcp1-3

(Neu5Aca2-6Galp1 -4GlcNAcp1 -2Mana1 )₂ -

93 Gala1-3Galp- 3,6-Manp1 -4GlcNAcp1 -4GlcNAcp-

(Galpl -4GlcNAcp1 -2Mana1 )₂-3,6-Manp1 -

94 Galp1-4Glcp- 4GlcNAcp1-4GlcNAcp-

(GlcNAcpl -2Mana1 )₂-3,6-Manp1 -

95 GalNAcp1-3Galp- 4GlcNAcp1-4GlcNAcp-

Fucal -2Galp1 -4(Fuca1 -3)GlcNAcp- 96 Gala-

Galp1-3(Fuca1-4) GlcNAcp- 97 GlcNAcp1-3Manp-

Neu5Aca- 98 Galp1-3GalNAca-

Neu5Aca2-3Galp1-3-(6-0-Su)GalNAca- 99 Gala1-3GalNAca-

(Galp1-4GlcNAc-p1-3)₃p- 100 Neu5Gca2-6GalNAca-

Mana1-6Manp- 101 Gala1-4Galp1-4GlcNAcp-

Neu5Aca2-6Galp1-4-(6-0-Su)GlcNAcp- 102 GalNAcal -3(Fuca1 -2)Gal-

Neu5Aca2-3Galp1-4-(6-0-Su)GlcNAcp- 103 Glcp- Structure of Glycan Structure of Glycan

Fuca1-3

GlcNAcp- 104 Glcp1-4Glcp- Neu5Aca2-3(6-0-Su)Galp1 -4

Neu5Aca2-8Neu5Aca-OCH₂C6H₄-p- 105 GlcAp1-6Galp-

3-0-Su-Galp1 -3(Fuca1 -4)GlcNAcp- 106 GalNAcp-

Glca- 107 GalNAcp1-4GlcNAcp-

3-O-Su-Galp- 108 Glca1-4Glcp-

GlcN(Gc)p- 109 Gala1-3GalNAcp-

Galp1-4GlcNAcp- 110 Neu5Aca2-6GalNAca-

Neu5Aca2-3Galp1 -4GlcNAcp- 111 Galp1-4(6-0-Su)Glcp-

Neu5Aca2-3Galp1 -3GalNAca- 112 Galp1-3Galp-

GlcNAcp1-3

Galp1-4GlcNAcp- 113 GalNAca- Galp1-4GlcNAcp1-6

Neu5Aca2-3Galp- 114 Gala1-6Glcp-

Manal -3(Mana1 -6)Manp- 115 Manp1-4GlcNAcp-

Neu5Aca2-3Galp1 -4Glcp- 116 Glcp1-6Glcp-

Galpl -4GlcNAcp1 -3Galp1 -4GlcNAcp- 117 GlcNAcpl -4GlcNAcp-Asn

3-0-Su-Galp1 -4GlcNAcp1 -3Galp1 -

118 Gala1-3Galp1-4GlcNAcp- 4GlcNAc-

Mana- 119 Fuca1-2Galp1-4Glcp-

Fucal -4GlcNAcp1 -3Galp1 -4Glcp-Fuca1 -

3-0-Su-Galp1-4Glcp- 120

2Galp1-3

Neu5Gca- 121 Galp1-2Galp-

(Glca1-4)₃p- 122 Gala1-4Galp1-4Glcp-

GlcNAcpl -3(GlcNAcp1 -6)Galp1 -4GlcNAcp- 123 Galp-

3-0-Su-Galp1-4GlcNAcp- 124 Galp1-3Galp1-4GlcNAcp-

GlcNAcpl -3Galp1 -4GlcNAcp- 125 Gala1-2Galp-

(Glca1-6)₄p- 126 6-O-Su-GlcNAcp-

GlcAp- 127 GlcNAcp1-4GlcNAcp-

Neu5Aca2-6Galp1 -4Glcp- 128 Galpl -4GlcNAcp1 -6GalNAca-

(Neu5Aca2-8)₃p- 129 GlcAp1-3GlcNAcp-

Fucal -2Galp1 -3(Fuca1 -4)GlcNAcp- 130 Galpl -4GlcNAcp1 -3Galp1 -4Glcp- Structure of Glycan Structure of Glycan

Fucal -3GlcNAcp1 -3Galp-Neu5Aca2- Fucal -3GlcNAcp1 -3Galp1 -4Glcp-Fuca1 -

131

3Galp1 -4 2Galp1 -4

6-H₂P0₃Mana- 132 Galp1 -3GalNAcp-

3-0-Su-Galp1 -4(6-0-Su)GlcNAcp- 133 GlcNAcp1 -3GalNAca-

Galpl -4GlcNAcp1 -6Galp1 -4GlcNAcp - 134 Gala1 -3(Fuca1 -2)Galp-

3,6-0-Su₂-Galp1 -4GlcNAcp- 135 GalNAcp1 -4Galp1 -4Glcp-

Glca1 -6Glca1 -6Glcp- 136 GlcNAcpl -GalNAca-

Galpl -4GlcNAcp1 -6(Galp1 -3)GalNAca- 137 Galpl -3GlcNAcp1 -3Galp1 -4Glcp-

Neu5Aca-OCH2C₆H₄-p- 138 GalNAcal -3(Fuca1 -2)Galp-

6-0-Su-Galp1 -4GlcNAcp- 139 GlcNAcpl -4-(Mur)-3-0-GlcNAcp-

Neu5Aca2-3Galp1 -3(Fuca1 -4)GlcNAcpl- 140 GlcNAcp-

GlcAa- 141 Galp1 -3GlcNAcp-

Galp1 -4(6-0-Su)GlcNAcp- 142 GalNAca1 -3Galp-

Neu5Aca2-8Neu5Aca2-3Galp1 -4Glcp- 143 GlcNAcpl -3(GlcNAcp1 -6)GalNAca-

Fuca- 144 Galal -3(Fuca1 -2)Galp1 -4GlcNAcp-

3-0-Su-Galp1 -3GalNAca- 145 Galpl -3GalNAcp1 -4Galp1 -4Glcp-

6-0-Su-Galp1 -4(6-0-Su)GlcNAcp- 146 Gala1 -4GlcNAcp-

3,6-0-Su2-Galp1 -4(6-0-Su)GlcNAcp- 147 Fuca1 -4GlcNAcp-

3-0-Su-Galp1 -4(6-0-Su)Glcp- 148 Fuca1 -3GlcNAcp-

6-0-Su-Galp1 -4(6-0-Su)Glcp- 149 Rhaa-

Neu5Aca2-6(Galp1 -3)GalNAca- 150 Gala1 -3(Fuca1 -2)Galp-

(Glca1 -4)₄p- 151 GlcNAcpl -4GlcNAcp1 -4GlcNAcp-

Neu5Aca2-6(Galp1 -3)GlcNAcp1 -3Galp1 -

152 4-0-Su-Galp1 -4GlcNAcp- 4Glcp-

Galpl -4GlcNAcp1 -3GalNAca-Galp1 -

153 GalNAca1 -3GalNAcp- 4GlcNAcp1 -6

Fucal -2Galp1 -3GlcNAcp- 154 (GlcNAcp1 -4)₆p-

Fucal -2Galp1 -3GalNAca- 155 Neu5Aca2-3Galp1 -3GlcNAcp-

6-H2PO3GICP- 156 (GlcNAcp1 -4)₅p-

6-0-Su-Galp1 -4Glcp- 157 GlcNAcpl -4Mur-L-Ala-D-i-Gln-Lys

Galal -3Galp1 -4(Fuca1 -3)GlcNAcp- 158 3-0-Su-Galp1 -3GlcNAcp- Parameters that describe properties of immunoprofiles of carbohydrate-binding antibodies are median and median absolute deviation. A low median indicates that most donors show low intensities of antibodies bound to the given glycan, while a large median suggests that there is a significant number of donors with large antibody binding intensities to the given glycan. Any of the glycan moieties in Tables 2A and 2B may be used as the monosaccharide or oligosaccharide moiety herein.

In some embodiments, a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) may contain one or more monosaccharide or oligosaccharide moieties having the structures

V. Antibacterial Agents

In some embodiments, one or more antibacterial agents may be administered in combination (e.g., administered substantially simultaneously (e.g., in the same pharmaceutical composition or in separate pharmaceutical compositions) or administered separately at different times) with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)).

Antibacterial agents may be grouped into several classes, e.g., quinolones, carbapenems, macrolides, DHFR inhibitors, aminoglycosides, ansamycins (e.g., geldanamycin, herimycin, and rifaximin), carbacephem (e.g., loracarbef), cephalosporins (e.g., cefadroxil, cefaolin, cefalotin, cefalothin, cephalexin, e.g., cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime, cefdinir, cefditoren,

cefoperazone, cefotaxime, cefpodoxime, ceftazidime, and ceftobiprole), glycopeptides (e.g., teicoplanin, vancomycin, telavancin, dalbavancin, and oritavancin), lincosamides (e.g., clindamycin and lincomycin), lipopeptides (e.g., daptomycin), monobactams (e.g., aztreonam), nitrofurans (e.g., furazolidone and nitrofurantoin), oxazolidinones, pleuromutilins, penicillins , sulfonamides, and tetracyclines (e.g., eravacycline, demeclocycline, doxycycline, minocycline, oxytetracycline, and tetracycline). Quinolones include, but are not limited to, ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, nalidixic acid, norfloxacin, ofloxacin, trovafloxacin, grepafloxacin, sparfloxacin, and temafloxacin. Carbapenems include, but are not limited to, ertapenem, doripenem,

imipenem/cilastatin, and meropenem. Macrolides include, but are not limited to, solithromycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, and spiramycin. In some embodiments, a macrolide is solithromycin. DHFR inhibitors include, but are not limited to, diaminoquinazoline, diaminopyrroloquinazoline, diaminopyrimidine, diaminopteridine, and diaminotriazines. Aminoglycosides include, but are not limited to, plazomicin, amikacin, gentamicin, gamithromycin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, streptomycin, and spectinomycin. Oxazolidinones include, but are not limited to, linezolid, tedizolid, posizolid, radezolid, and furazolidone. Pleuromutilins include, but are not limited to, retapamulin, valnemulin, tiamulin, azamulin, and lefamulin. Penicillins include, but are not limited to, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, penicillin G, temocillin, and ticarcillin. Sulfonamides include, but are not limited to, mafenide, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfadimethoxine, sulfamethizole, sulfamethoxazole, sulfanamide, sulfasalazine, sulfisoxazole, trimethoprim-sulfamethoxazole (Co-trimoxazole) (TMP-SMX), and sulfonamidochrysoidine.

In some embodiments, the antibacterial agent used in combination with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) is selected from the group consisting of Iinezolid, tedizolid, posizolid, radezolid, retapamulin, valnemulin, tiamulin, azamulin, lefamulin, plazomicin, amikacin, gentamicin, gamithromycin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, streptomycin, spectinomycin, geldanamycin, herbimycin, rifaximin, loracarbef, ertapenem, doripenem, imipenem/cilastatin, meropenem, cefadroxil, cefazolin, cefalotin, cefalexin, cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepime, ceftaroline fosamil, ceftobiprole, teicoplanin, vancomycin, telavancin, dalbavancin, oritavancin, clindamycin, lincomycin, daptomycin, solithromycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin, aztreonam, furazolidone, nitrofurantoin, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin g, penicillin v, piperacillin, penicillin g, temocillin, ticarcillin, amoxicillin clavulanate, ampicillin/sulbactam,

In some embodiments, the antibacterial agent used in combination with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) is tedizolid, azithromycin, meropenem, amikacin, levofloxacin, rifampicin, Iinezolid, erythromycin, or solithromycin. In some embodiments, the antibacterial agent used in combination with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) is tedizolid, azithromycin, meropenem, amikacin, or levofloxacin. In some embodiments, the antibacterial agent used in combination with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) is tiamulin. In some embodiments, the antibacterial agent used in combination with a compound described herein (e.g., a compound of any one of formulas (I)- (XXXXX)) is solithromycin. VI. Methods

Methods described herein include, e.g., methods of protecting against or treating a bacterial infection (e.g., a Gram-negative bacterial infection) in a subject and methods of preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria (e.g., Gram-negative bacteria). A method of treating a bacterial infection (e.g., a Gram-negative bacterial infection) in a subject includes administering to the subject a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) or a pharmaceutical composition thereof. In some embodiments, the bacterial infection is caused by Gram- negative bacteria. In some embodiments, the bacterial infection is caused by a resistant strain of bacteria. In some embodiments, the resistant strain of bacteria is a resistant strain of E. coli. A method of preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria (e.g., Gram-negative bacteria) includes contacting the bacteria (e.g., Gram-negative bacteria) or a site susceptible to bacterial growth (e.g., Gram-negative bacterial growth) with a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) or a pharmaceutical composition thereof. In some embodiments, the bacterial infection is caused by Gram-negative bacteria. In some embodiments, the bacterial infection is caused by a resistant strain of bacteria. In some embodiments, the resistant strain of bacteria is a resistant strain of E. coli. In some embodiments, a compound used in any methods described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) may bind to LPS in the cell membrane of Gram- negative bacteria to disrupt and permeabilize the cell membrane, leading to cell death and/or sensitization to other antibiotics. Additionally, the monosaccharide or oligosaccharide moieties in the compounds described herein also serve as a gradient against which immune cells chemotax to the site of bacterial infection and/or growth.

Moreover, methods described herein also include methods of protecting against or treating sepsis in a subject by administering to the subject a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)). In some embodiments, the method further includes administering to the subject an antibacterial agent. Methods described herein also include methods of preventing lipopolysaccharides (LPS) in Gram-negative bacteria (e.g., a resistant strain of Gram-negative bacteria or a resistant strain of E. coli (e.g., E. coli BAA-2469)) from activating a immune system in a subject by administering to the subject a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)). In some embodiments of the method, the method prevents LPS from activating a macrophage. In some embodiments, the method further includes administering to the subject an antibacterial agent. In some embodiments, a compound used in any methods described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) may bind to LPS in the cell membrane of Gram-negative bacteria to disrupt and permeabilize the cell membrane, leading to cell death and/or sensitization to other antibiotics.

In some embodiments, the methods described herein may further include administering to the subject an antibacterial agent in addition to a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)). Methods described herein also include methods of protecting against or treating a bacterial infection in a subject by administering to said subject (1 ) a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) and (2) an antibacterial agent. Methods described herein also include methods of preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria, by contacting the bacteria or a site susceptible to bacterial growth with (1 ) a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) and (2) an antibacterial agent.

In some embodiments, the compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) is administered first, followed by administering of the antibacterial agent alone. In some embodiments, the antibacterial agent is administered first, followed by administering of the compound described herein alone. In some embodiments, the compound described herein and the antibacterial agent are administered substantially simultaneously (e.g., in the same pharmaceutical composition or in separate pharmaceutical compositions). In some embodiments, the compound described herein or the antibacterial agent is administered first, followed by administering of the compound described herein and the antibacterial agent substantially simultaneously (e.g., in the same pharmaceutical composition or in separate pharmaceutical compositions). In some embodiments, the compound described herein and the antibacterial agent are administered first substantially simultaneously (e.g., in the same pharmaceutical composition or in separate pharmaceutical compositions), followed by administering of the compound described herein or the antibacterial agent alone. In some embodiments, when a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) and an antibacterial agent are administered together (e.g., substantially simultaneously in the same or separate pharmaceutical compositions, or separately in the same treatment regimen), the MIC of each of the compound and the antibacterial agent may be lower than the MIC of each of the compound and the antibacterial agent when each is used alone in a treatment regimen.

VII. Pharmaceutical Compositions and Preparations

A compound described herein may be formulated in a pharmaceutical composition for use in the methods described herein. In some embodiments, a compound described herein may be formulated in a pharmaceutical composition alone. In some embodiments, a compound described herein may be formulated in combination with an antibacterial agent in a pharmaceutical composition. In some embodiments, the pharmaceutical composition includes a compound described herein (e.g., a compound described by any one of formulas (l)-(XXXXX)) and pharmaceutically acceptable carriers and excipients.

Acceptable carriers and excipients in the pharmaceutical compositions are nontoxic to recipients at the dosages and concentrations employed. Acceptable carriers and excipients may include buffers such as phosphate, citrate, HEPES, and TAE, antioxidants such as ascorbic acid and methionine, preservatives such as hexamethonium chloride, octadecyldimethylbenzyl ammonium chloride, resorcinol, and benzalkonium chloride, proteins such as human serum albumin, gelatin, dextran, and

immunoglobulins, hydrophilic polymers such as polyvinylpyrrolidone, amino acid residues such as glycine, glutamine, histidine, and lysine, and carbohydrates such as glucose, mannose, sucrose, and sorbitol.

Examples of other excipients include, but are not limited to, antiadherents, binders, coatings, compression aids, disintegrants, dyes, emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, sorbents, suspensing or dispersing agents, or sweeteners. Exemplary excipients include, but are not limited to: butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine,

methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

The compounds herein may have ionizable groups so as to be capable of preparation as pharmaceutically acceptable salts. These salts may be acid addition salts involving inorganic or organic acids or the salts may, in the case of acidic forms of the compounds herein be prepared from inorganic or organic bases. Frequently, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well-known in the art, such as hydrochloric, sulphuric, hydrobromic, acetic, lactic, citric, or tartaric acids for forming acid addition salts, and potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, various amines, and the like for forming basic salts. Methods for preparation of the appropriate salts are well-established in the art.

Representative acid addition salts include, but are not limited to, acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate,

camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and valerate salts. Representative alkali or alkaline earth metal salts include, but are not limited to, sodium, lithium, potassium, calcium, and magnesium, as well as nontoxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine.

Depending on the route of administration and the dosage, a compound herein or a

pharmaceutical composition thereof used in the methods described herein will be formulated into suitable pharmaceutical compositions to permit facile delivery. A compound (e.g., a compound of any one of formulas (l)-(XXXXX)) or a pharmaceutical composition thereof may be formulated to be administered intramuscularly, intravenously (e.g., as a sterile solution and in a solvent system suitable for intravenous use), intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctival, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularally, orally (e.g., a tablet, capsule, caplet, gelcap, or syrup), topically (e.g., as a cream, gel, lotion, or ointment), locally, by inhalation, by injection, or by infusion (e.g., continuous infusion, localized perfusion bathing target cells directly, catheter, lavage, in cremes, or lipid compositions). Depending on the route of administration, a compound herein or a pharmaceutical composition thereof may be in the form of, e.g., tablets, capsules, pills, powders, granulates, suspensions, emulsions, solutions, gels including hydrogels, pastes, ointments, creams, plasters, drenches, osmotic delivery devices, suppositories, enemas, injectables, implants, sprays, preparations suitable for iontophoretic delivery, or aerosols. The compositions may be formulated according to conventional pharmaceutical practice.

A compound described herein may be formulated in a variety of ways that are known in the art.

For use as treatment of human and animal subjects, a compound described herein can be formulated as pharmaceutical or veterinary compositions. Depending on the subject (e.g., a human) to be treated, the mode of administration, and the type of treatment desired, e.g., prophylaxis or therapy, a compound described herein is formulated in ways consonant with these parameters. A summary of such techniques is found in Remington: The Science and Practice of Pharmacy, 22nd Edition, Lippincott Williams & Wilkins (2012); and Encyclopedia of Pharmaceutical Technology, 4th Edition, J. Swarbrick and J. C. Boylan, Marcel Dekker, New York (2013), each of which is incorporated herein by reference.

Formulations may be prepared in a manner suitable for systemic administration or topical or local administration. Systemic formulations include those designed for injection (e.g., intramuscular, intravenous or subcutaneous injection) or may be prepared for transdermal, transmucosal, or oral administration. The formulation will generally include a diluent as well as, in some cases, adjuvants, buffers, and preservatives. The compounds can be administered also in liposomal compositions or as microemulsions. Systemic administration may also include relatively noninvasive methods such as the use of suppositories, transdermal patches, transmucosal delivery and intranasal administration. Oral administration is also suitable for compounds herein. Suitable forms include syrups, capsules, and tablets, as is understood in the art.

The pharmaceutical compositions can be administered parenterally in the form of an injectable formulation. Pharmaceutical compositions for injection can be formulated using a sterile solution or any pharmaceutically acceptable liquid as a vehicle. Formulations may be prepared as solid forms suitable for solution or suspension in liquid prior to injection or as emulsions. Pharmaceutically acceptable vehicles include, but are not limited to, sterile water, physiological saline, and cell culture media (e.g., Dulbecco's Modified Eagle Medium (DMEM), a-Modified Eagles Medium (a-MEM), F-12 medium). Such injectable compositions may also contain amounts of nontoxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, such as sodium acetate and sorbitan monolaurate. Formulation methods are known in the art, see e.g., Pharmaceutical Preformulation and Formulation, 2nd Edition, M. Gibson, Taylor & Francis Group, CRC Press (2009).

The pharmaceutical compositions can be prepared in the form of an oral formulation.

Formulations for oral use include tablets containing the active ingredient(s) in a mixture with non-toxic pharmaceutically acceptable excipients. These excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystalline cellulose, starches including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulating and disintegrating agents (e.g., cellulose derivatives including microcrystalline cellulose, starches including potato starch, croscarmellose sodium, alginates, or alginic acid); binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, carboxymethylcellulose sodium, methylcellulose, hydroxypropyl methylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricating agents, glidants, and antiadhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenated vegetable oils, or talc). Formulations for oral use may also be provided as chewable tablets, or as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin), or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil. Powders, granulates, and pellets may be prepared using the ingredients mentioned above under tablets and capsules in a conventional manner using, e.g., a mixer, a fluid bed apparatus or a spray drying equipment.

Other pharmaceutically acceptable excipients for oral formulations include, but are not limited to, colorants, flavoring agents, plasticizers, humectants, and buffering agents. Formulations for oral use may also be provided as chewable tablets, or as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate or kaolin), or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil. Powders, granulates, and pellets may be prepared using the ingredients mentioned above under tablets and capsules in a conventional manner using, e.g., a mixer, a fluid bed apparatus or a spray drying equipment.

Dissolution or diffusion controlled release of a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) or a pharmaceutical composition thereof can be achieved by appropriate coating of a tablet, capsule, pellet, or granulate formulation of the compound, or by incorporating the compound into an appropriate matrix. A controlled release coating may include one or more of the coating substances mentioned above and/or, e.g., shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethylcellulose, acrylic resins, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinyl pyrrolidone, polyethylene, polymethacrylate, methylmethacrylate, 2-hydroxymethacrylate, methacrylate hydrogels, 1 ,3 butylene glycol, ethylene glycol methacrylate, and/or polyethylene glycols. In a controlled release matrix formulation, the matrix material may also include, e.g., hydrated

methylcellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and/or halogenated fluorocarbon.

The pharmaceutical composition may be formed in a unit dose form as needed. The amount of active component, e.g., a compound described herein (e.g., a compound of any one of formulas (I)- (XXXXX)), included in the pharmaceutical compositions are such that a suitable dose within the designated range is provided (e.g., a dose within the range of 0.01 -100 mg/kg of body weight). VIII. Routes of Administration and Dosages

In any of the methods described herein, compounds herein may be administered by any appropriate route for treating or protecting against a bacterial infection (e.g., a Gram-negative bacterial infection), or for preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria (e.g., Gram- negative bacteria). Compounds described herein may be administered to humans, domestic pets, livestock, or other animals with a pharmaceutically acceptable diluent, carrier, or excipient. In some embodiments, administering comprises administration of any of the compounds described herein (e.g., compounds of any one of formulas (l)-(XXXXX)) or compositions intramuscularly, intravenously (e.g., as a sterile solution and in a solvent system suitable for intravenous use), intradermal^, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleurally,

intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctival, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularally, orally (e.g., a tablet, capsule, caplet, gelcap, or syrup), topically (e.g., as a cream, gel, lotion, or ointment), locally, by inhalation, by injection, or by infusion (e.g., continuous infusion, localized perfusion bathing target cells directly, catheter, lavage, in cremes, or lipid compositions). In some embodiments, if an antibacterial agent is also administered in addition to a compound described herein, the antibacterial agent or a pharmaceutical composition thereof may also be administered in any of the routes of administration described herein.

The dosage of a compound described herein (e.g., a compound of any one of formulas (I)- (XXXXX)) or a pharmaceutical compositions thereof depends on factors including the route of administration, the disease to be treated (e.g., the extent and/or condition of the bacterial infection), and physical characteristics, e.g., age, weight, general health, of the subject. Typically, the amount of the compound or the pharmaceutical composition thereof contained within a single dose may be an amount that effectively prevents, delays, or treats the bacterial infection without inducing significant toxicity. A pharmaceutical composition may include a dosage of a compound described herein ranging from 0.01 to 500 mg/kg (e.g., 0.01 , 0.1 , 0.2, 0.3, 0.4, 0.5, 1 , 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg/kg) and, in a more specific embodiment, about 0.1 to about 30 mg/kg and, in a more specific embodiment, about 1 to about 30 mg/kg. In some embodiments, when a compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) and an antibacterial agent are administered together (e.g., substantially simultaneously in the same or separate

pharmaceutical compositions, or separately in the same treatment regimen), the dosage needed of the compound described herein may be lower than the dosage needed of the compound if the compound was used alone in a treatment regimen.

A compound described herein (e.g., a compound of any one of formulas (l)-(XXXXX)) or a pharmaceutical composition thereof may be administered to a subject in need thereof, for example, one or more times (e.g., 1 -10 times or more; 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 1 0 times) daily, weekly, monthly, biannually, annually, or as medically necessary. Dosages may be provided in either a single or multiple dosage regimens. The timing between administrations may decrease as the medical condition improves or increase as the health of the patient declines. The dosage and frequency of administration may be adapted by the physician in accordance with conventional factors such as the extent of the infection and different parameters of the subject.

EXAMPLES

General Schemes

The compounds of the disclosure and their precursors may be synthesized by a number of methods known to those skilled in the art. The following Scheme 1 and Scheme 2 illustrate several methods and strategies that may be employed but are not meant to be limiting. Cyclic heptapeptides that comprise residues M1 and M2 may be prepared by synthesis using solid phase peptide methods or by solution phase methods. Alternately, the cyclic heptapeptides that comprise residues M1 and M2 may be derived from certain compounds known as polymyxins or from octapeptins which may be derived from fermentation sources or from synthetic sources. The protected M1 and M2 residues may be obtained by first protection of the polymyxin or octapeptin followed by enzymatic hydrolysis (e.g. with savinase or other enzymes known to those skilled in the art). Synthetic, non-natural polymyxins or their

corresponding cyclic heptapeptides, may also be used in the preparation of M1 and M2 and may be prepared by methods known to those who are skilled in the art.

The cyclic heptapeptides may be optionally derivatized with 1 , 2 or 3 amino acids which may be the same or different. Using a suitable linker group L, the two halves which may be the same or different, may be coupled. Appending a suitable E group, which may or may not contain part of the linker L, and subsequent deprotection affords the compounds of the present disclosure.

Alternatively, the linker group L' may be prepared from the individual amino acid groups and L by methods known to those in the art. L' may be built up sequentially or by a convergent synthesis and then coupled to the cyclic heptapeptides that comprise the residues M1 and M2. The L' group may be assembled with one or more E groups attached or the E group(s) may be appended after complete construction of L'. After deprotection, the compounds of the immediate disclosure may be obtained. Alternatively, when a, b, a', and b' are 1 , A1 -M1 or A2-M2 may be derived from a polymyxin or octapeptin precursor by first protection followed by enzymatic hydrolysis (e.g. with papain). Subsequent coupling with L, E or L'-E groups and deprotection, if required, provides the compounds of the present disclosure. Alternatively, when a, b, c, a', b' and c' are 0, the cyclic heptapeptides that comprise the residues M1 and M2 may be coupled directly with L, E or L'-E groups followed by deprotection, if required, to give the compounds of the immediate disclosure.

Other methods known by those skilled in the art may be employed to prepare symmetric compounds (i.e. those where A1 and A2 are the same and M1 and M2 are the same), or asymmetric compounds where A1 -M1 is different from A2-M2 and where L may or may not be symmetric.

The E group(s) may be appended at the time L' is synthesized or it may be appended after the L' is completely assembled or it may be appended after M1 , M2 and L' are coupled. Using orthogonal protection schemes known to those skilled in the art, allows different E groups (e.g. a-L-Rha, a-Gal epitope, laminarin) to be attached in the same molecule. The E groups themselves may possess part of the final linker group L' and as such, L' would be made up of the linker used to assemble M2-L'-M1 and

(E)n

E-L' to give the compounds of the present disclosure ^ . Scheme 1. Synthesis of compound

E

M2— L'— M

Scheme 2. Synthesis of compounds

General Methods

Preparative HPLC was performed using the following: Teledyne Isco HP C1 8, 50g column. Eluent: CH3CN/H2O/ 0.1 % formic acid or 0.1 % trifluoroacetic acid; various linear gradients as necessary at 40 mL /min on an Isco Combiflash Rf LC unit, or Isco EZ prep HPLC, with UV Detection at 220 and 254 nm on a Luna 5 micron C1 8, 100 A, AXIA 1 00 x 30 mm. Eluent: CH3CN/H2O/ 0.1 % formic acid or 0.1 % trifluoroacetic acid; various linear gradients as necessary at 25 mL /min on the Gilson System ; 215 Liquid Handler, Gilson UV-VIS 155, Gilson 305 pump and Detector. UV detection at 220 and 254 nm.

Analytical LC/MS: High resolution liquid chromatography mass spectrometry (HRES-LC/MS) was performed using a Waters Q-TOF Premier mass spectrometer with an electrospray probe coupled with a Waters Acquity H-class UPLC system with a diode array detector set to collect from 210 nm to 400 nm. A gradient of 0.1 % formic acid in water (A) and 0.1 % formic acid in acetonitrile (B) was run from 15%B to 95% over 10 min using a 100 x 2.1 mm 1 .7 μ Phenomenex Kinetex C18 column at 40 °C.

Liquid chromatography mass spectrometry was performed using an Agilent 6120 mass spectrometer an electrospray probe coupled with an Agilent 1260 HPLC system with a variable wavelength detector set to either 220 nm or 254 nm. A gradient of 0.1 % formic acid in water (A) and 0.1 % formic acid in acetonitrile (B) was run from 15%B to 99% over 3.5 min using a 50x3.0 mm 2.6μ Phenomenex Gemini-NX column at 30°C.

1 H-NMR (1 H NMR) spectra were acquired on a Bruker 300 MHz system using a 5 mm QNP probe and chemical shifts are reported as ppm (δ) downfield from tetramethylsilane.

Compounds herein may be made using synthetic methods known in the art, including procedures analogous to those disclosed below. Exemplary protocols, techniques, reagents, and solvents for synthetic methods known in the art are provided, for example, in Caren, S., Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques. Wiley, 201 1 , and Benoiton, N.L., Chemistry of Peptide Synthesis. CRC Press, 2005, each of which is incorporated by reference in their entirety.

Example 1. Synthesis of L-Rhamnose-PEG1-NH2 (INT-1 )

Step a. Synthesis of benzyl (2-(2-hydroxyethoxy)ethyl)carbamate

To a solution of 2-(2-aminoethoxy)ethanol (10.5 g, 0.1 mol) in 200 mL THF was added Hunig's base (28 mL, 0.2mol) under the ice-water bath, then CBz-CI (18g, 0.1 mol) in 50 mL THF was added dropwise to the above cooled solution. The resultant solution was stirred for overnight and then concentrated to remove solvent, the residue was partitioned by ethyl acetate (300mL) and water 150 mL, the organic layer was washed by 5% sodium bicarbonate (100 mL), brine (1 00 mL), and 1 N HCI (1 00 mL) and then brine, respectively. The organic layer was dried and concentrated for next step. Yield of products 18 g, 75%. Ή NMR (300 MHz, DMSO-afe) δ 7.34 (d, J = 3.2 Hz, 5H), 5.01 (d, J = 2.5 Hz, 2H), 3.40 (m, 8H).

Step b. Synthesis of (2R,3R,4R,5S,6S)-2-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethoxy)-6- methyltetrahydro-2H-pyran-3,4,5-triyl triacetate

To a solution of benzyl (2-(2-hydroxyethoxy)ethyl)carbamate (14g, 56 mmol) and per-acetyl-L- rhamnose (20 g, 56 mmol) in 200 mL dry DCM under the ice-water bath, then BF₃-Et20 (15 mL, 1 10 mmol) was added dropwise to the above cooled solution. After stirring overnight, the reaction was quenched with saturated NaHCCb (aq) and diluted with EtOAc. The organic phase was washed sat. NaHC03 (aq) and brine. The combined organic layers were dried over anhydrous Na2S04. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 1 00% acetonitrile and water, using 0.1 %

trifluoroacetic acid as the modifier. The product was isolated as an oil, 18 g (63%). LC/MS [M]+1 512.2.

Step c. Synthesis of benzyl (2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)ethoxy)ethyl)carbamate

A solution of (2R,3R,4R,5S,6S)-2-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethoxy)-6- methyltetrahydro-2H-pyran-3,4,5-triyl triacetate (26 g, 50 mmol) and hydrazine (25 mL) in anhydrous methanol was stirred overnight, The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, 18.5 g (95%). LC/MS [M+H]⁺ 386.2. The material was subjected to high vacuum to remove excess hydrazine.

Step d. Synthesis of (2R,3R,4R,5R,6S)-2-(2-(2-aminoethoxy)ethoxy)-6-methyltetrahydro-2H-pyran- 3,4,5-triol (L-Rhamnose-PEG1-NH2, INT-1)

Benzyl (2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)ethoxy)ethyl)carbamate (14.6 mg, 38 mmol) was dissolved into 30 mL MeOH and 30 mL ethyl acetate, then 5 mg of 5% Pd on charcoal was added above solution, and the mixture was stirred at room temperature under a hydrogen atmosphere overnight. The palladium on charcoal was removed by filtration after completion of the reaction by LCMS. The filtrate was concentrated and used in the next step without any purification. ¹ H NMR (300 MHz, methanol-dt) δ 4.75(s, 1 H) 3.84 (ddt, J = 8.5, 3.3, 1 .6 Hz,

2H), 3.76 - 3.52 (m, 7H), 3.47 - 3.28 (m, 4H), 3.15 (t, J = 5.1 Hz, 2H), 1 .28 (d, J = 6.2 Hz, 3H). ¹³C NMR (75 MHz, methanol-ck) δ 100.36, 72.52, 70.93, 70.72, 70.1 1 , 68.45, 66.60, 66.27, 39.29, 16.61 . LCMS 252.2 [M+H]⁺ Example 2. Synthesis of 4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoic acid (INT-2)

Procedu e A

Succinic anhydride (156 mg, 1 .57 mmol) was added to a stirring mixture of INT-1 (375 mg, 1 .49 mmol) and DIEA (193 mg, 1 .49 mmol) in MeOH (7 mL) and the reaction was stirred at RT for 3 hours at room temperature. The solvent was reduced by 80% on the rotary evaporator and the mixture was applied to reversed phase HPLC (5 to 50% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and lyophilized to afford 4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid (INT-2) as a clear vicous oil. Yield: 75%. LC/MS [M-H-]- 350.2. Procedu e B

Step a. Synthesis of benzyl (2-(2-hydroxyethoxy)ethyl)carbamate

To the solution of 2-(2-aminoethoxy)ethanol (301 .0 g, 2.863mol, 1 .Oeq) in 1500ml_ DCM was added Et₃N (343. Og, 3.390mol, 1 .2eq) under the ice-water bath (control the temperature below 20°C), then CBz-CI (488. Og, 2.863mol, 1 .Oeq) was added dropwise to the above cooled solution (control inside temperature below 20°C). The resultant solution was stirred for overnight (>16 hrs) and then washed with water 1500 mL, sat. NaHC0₃ (1500 mL), 1 N HCI (1500 mL) and brine (1500 mL), respectively. The organic layer was dried (200 g of Na2S04) and concentrated for next step directly. Yield of product 639 g, 92%. Colorless oil.

Step b. Per-acetylation of L-rhamnose

To a solution of L-rhamnose (69.0 g, 0.3799 mol, 1 .0 eq) and DMAP (1 .40 g, 0.02x) in pyridine (280 mL, 4V/M) was added acetic anhydride (232.0 g, 2.273 mol, 6.0 equiv) dropwise under the ice-water bath (to control the temperature of the reaction below 20°C), The mixture was stirred at ambient temperature (>12 hrs), and volatiles were removed under reduced pressure (toluene azeotrope). The crude product was dissolved in EtOAc (800 mL), washed with hydrochloric acid (1 N, 800 mL^*2), sat. NaHCO3 (800 mL), brine (800 mL), and dried with sodium sulfate (100 g). Volatiles were removed under reduced pressure, and crude per-acetyl-L-rhamnose (131 .0 g, yield: 100%) was used directly for the next step. Step c. Synthesis of (2R,3R,4R,5S,6S)-2-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethoxy)-6- methyltetrahydro-2H-pyran-3,4,5-triyl triacetate

To the solution of benzyl (2-(2-hydroxyethoxy)ethyl)carbamate (48.8 g, 0.204 mol, 1 .20 eq) and per-acetyl-L-rhamnose (56.6 g, 0.170 mol, 1 .00 eq) in 500 mL dry DCM under the ice-water bath, then BF3-Et20 (52.5 mL, 0.416 mol, 2.5 eq) was added dropwise to the above cooled solution (control the temperature below 20°C). After stirring overnight, the reaction was quenched by 700 mL of sat. NaHCCb (aq), concentrated and purified by chromatography on silica gel (90g/500g), eluent:PE (1 L),

PE/EA=5/1 (10.8L), PE/EA=3/1 (20L). Yield: 50.0 g, 57%, of a colorless oil.

Step d. Synthesis of benzyl (2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)ethoxy)ethyl)carbamate

To a solution of (2R,3R,4R,5S,6S)-2-(2-(2-(((benzyloxy)carbonyl)amino)ethoxy)ethoxy)-6- methyltetrahydro-2H-pyran-3,4,5-triyl triacetate (33.7 g, 0.066 mol, 1 .0 eq) in methanol (200 mL) was added NaOMe (1 .14 g in 50 mL of MeOH, 0.050 mol, 0.75 eq) and stirred overnight. The resultant solution was concentrated and dissolved in 800 mL of THF, then filtered through a pad of silica gel (20 g), and washed with THF (400 mL^*3) and concentrated to give a colorless oil (23.4 g, 92%). Step e. Synthesis of (2R,3R,4R,5R,6S)-2-(2-(2-aminoethoxy)ethoxy)-6-methyltetrahydro-2H-pyran- 3,4,5-triol (L-Rhamnose-PEG1-NH2) (INT-1)

Benzyl (2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)ethoxy)ethyl)carbamate (21 .2 g, 0.066 mol, 1 .0 eq) was dissolved into 300 mL MeOH, then 2.2 g of 5% palladium on charcoal (0.1 x) was added to the above solution and the mixture was stirred at room temperature under the hydrogen atmosphere for overnight. The palladium on charcoal was removed by filtration after the reaction was complete as judged by TLC. The filtrate was concentrated to give INT-1 as a colorless oil (13.4 g, 97%). LCMS 252.2[M+H]⁺ Step f. Synthesis of 4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoi -2)

A solution of INT-1 (10.93 g, 0.043 mol, 1 .0 eq) and succinic anhydride (4.35 g, 0.043 mol, 1 .0 eq) in anhydrous MeOH (1 00 mL) was stirred at RT for overnight (>12 hrs). Then the solvent was removed and the residue was purified by flash silica gel column chromatography (18 g/100 g), eluent: DCM/MeOH=10/1 (1 .8L), DCM/MeOH=6/1 (1 .6L) to give INT-2 (12.1 g, 82 %) as a colorless oil. ¹ HNMR (400 MHz, methanol-ck): 4.72 (s, 1 H); 3.82-3.75 (m, 2H); 3.67-3.53 (m, 7H); 3.39-3.35 (m, 2H); 3.31 (m, 1 H); 2.61 -2.47 (m, 4H); 1 .26 (d, 6.4 Hz, 3H).

Example 3. Synthesis of 2-aminoethyl 2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl-(1->3)-2,4,6-tri-

0-acetyl-3-thio-beta-D-glucopyranosyl-(1->3)-2,4,6-tri-0-acetyl-1 ,3-dithio-beta-D-glucopyranoside

(INT-3)

Step a. Synthesis of (3aR,5R,6R,6aR)-5-((R)-2,2-dimethyl-1 ,3-dioxolan-4-yl)-2,2- dimethyltetrahydro-2H-furo[2,3-d][1 ,3]dioxol-6-yl trifluoromethanesulfonate

Triflic anhydride (7.8 mL, 46.39 mmol) was added over 30 minutes to a cold (0°C) solution of the diacetone allofuranose (10.50 g, 40.34 mmol) in pyridine (100 mL). After stirring in the ice-bath for 1 hour, the reaction was diluted with ethyl acetate (250 mL) and the organic layer was washed with a mixture of saturated sodium bicarbonate and brine (50 mL + 50 mL), dried (Na2S04) and concentrated under reduced pressure to provide the crude triflate. The crude residue was purified using a normal phase flash column (0-10% EA/Hex) to give the title compound (14.15 g, 89%) as a colorless oil. 1 HNMR (300 MHz, Chloroform-d): 5.85 (d, 1 H); 4.93 (dd, 1 H); 4.79 (dd, 1 H); 4.21 (dd, 1 H); 4.1 1 -4.17 (dt, 1 H); 3.90-3.95 (dt, 1 H); 1 .61 (s, 3H); 1 .47 (s, 3H); 1 .41 (s, 3H); 0.3 (s, 3H).

Step b. Synthesis of Thio-linked Disaccharide: ((2R,3R,4S,5R,6S)-2-[(acetyloxy)methyl]-6- {[(3aR,5R,6S,6aS)-5-(2,2-dimethyl-1 ,3-dioxolan-4-yl)-2,2-dimethyltetrahydro-2H-furo[2,3- d][1 ,3]dioxol-6-yl]sulfanyl}oxane-3,4,5-triyl triacetate)

Sodium hydride was added to a solution of 2,3,4,6-tetra-0-acetyl-l-thio-p-D-glucopyranose (14.00 g, 38.42 mmol) in dry THF (450 mL in a 2000-mL RB Flask) at 0 °C. The suspension was stirred under nitrogen until hydrogen formation had ceased. To this solution, 1 ,7,10-trioxa-4,13-diazacyclopentadecane (Kryptofix 21 , 4,10-diaza-16-crown-5-ether, 1 .53 g, 9.92 mmol) and a solution of (3aR,5R,6R,6aR)-5-((R)- 2,2-dimethyl-1 ,3-dioxolan-4-yl)-2,2-dimethyltetrahydro-2H-furo[2,3-d][1 ,3]dioxol-6-yl

trifluoromethanesulfonate (13.09 g, 46.39 mmol) in THF (200 mL) was added via cannula transfer and the mixture was stirred for 2 h at 0 °C under nitrogen, then at room temperature for overnight. The reaction mixture was concentrated under reduced pressure. The residue was dissolved in DCM (400 mL), washed with water (200 mL), dried (Na2S04), and concentrated. Added 40 mL of EtOH to the residue to dissolve the foam followed by swirling the flask. The foam slowly turn to white solid. After standing still for 5 minutes, breaking down the solid cake and filtering and washing with 20% aq. EtOH (100 mL). After drying, 10.28 g of the title compound was obtained (44%). Check TLC of the mother liquor and some of the product remained in the mother liquor which was purified with normal phase flash column. 1 HNMR (300 MHz, Chloroform-d): 5.86 (d, H), 5.25 (dd, 1 H), 5.10 (m, 2H), 4.86 (d, H), 4.76 (d, H), 3.95-4.42 (m, 6H), 3.64-3.78 (m, H), 3.58 (d, H), 2.1 1 (s, 3H), 2.09 (s,3H), 2.06 (s, 3H), 2.03 (s, 3H), 1 .54 (s, 3H), 1 .45 (s, 3H), 1 .35 (s, 3H), 1 .35 (s, 3H) Step c. Synthesis of 1 ,2,4,6-tetra-0-acetyl-3-S-(2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl)-3- thio-D-glucopyranose

A solution of ((2R,3R,4S,5R,6S)-2-[(acetyloxy)methyl]-6-{[(3aR,5R,6S,6aS)-5-(2,2-dimethyl-1 ,3- dioxolan-4-yl)-2,2-dimethyltetrahydro-2H-furo[2,3-d][1 ,3]dioxol-6-yl]sulfanyl}oxane-3,4,5-triyl triacetate) (10.28 g) in 60% aq. AcOH/TFA (20:1 , 1 05 mL) was heated for 5 hours at 70 °C, then concentrated under reduced pressure and the residue dissolved in water and freeze-dried. Conventional acetylation of the resulting solid (1 :1 Ac20/pyridine, 200 mL) yielded the product. Rotovaped to remove most of Ac20/Py, diluted with DCM (400 mL) and washed with 0.5 N HCI 200 mL. 200 mL of DCM extracted the aqueous layer once. Combined DCM extract and dried with Na2S04. DCM was removed under reduced pressure. The residue was precipitated with 30% EA/Hex to give a white solid. Filtration and 30%EA/Hex wash and dry. Check TLC of mother liquor, the impurities and the desired product have almost equal amount.

Purification was not performed due to the small amount in the mother liquor. The precipitation gave peracetylated (1 -3>)-thiodisaccharide (1 ,2,4,6-tetra-0-acetyl-3-S-(2,3,4,6-tetra-0-acetyl-beta-D- glucopyranosyl)-3-thio-D-glucopyranose) 9.770 g (83.0%) and a column purification gave 1 .384 g (1 1 .8%). 1 HNMR showed alpha/beta ratio 1 :1 (from ~H NMR integration of H-1 signals).

Step d. Synthesis of (1->3)-Thiodisaccharide Bromide: 2,4,6-tri-0-acetyl-3-S-(2,3,4,6-tetra-0-acetyl- beta-D-g lucopyranosyl)-3-th io-alpha-D-g lucopyranosyl bromide

To a solution of 1 ,2,4,6-tetra-0-acetyl-3-S-(2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl)-3-thio- D-glucopyranose (1 1 .15 g, 16.06 mmol) in dry DCM (1 60 mL) at 0 °C was added dropwise commercial 33% HBr in AcOH (34.9 mL, 12 equiv.). After the mixture was stirred for 3 hours in an ice-bath, TLC (1 :1 EtOAc-hexane) showed complete conversion of the SM into a single product. DCM was added (240 mL), and the solution was washed with cold water (2 x 100 mL) and dried with Na2S04. DCM was removed under reduced pressure. The residue was precipitated with 30% EA/Hex to give a white solid. Filtration and 30% EA/Hex wash and dry to give the title compound (10.50 g, 91 .4%) as white a powder. 1 HNMR spectrum was identical with literature data for the compound [B. Sylla, et al.; J. Med.

Chem., 2014, 57 (20), pp 8280-8292].

Step e. Synthesis of (1->3)-Thiodisaccharide Thiol: 2,4,6-tri-0-acetyl-3-S-(2,3,4,6-tetra-0-acetyl- beta-D-glucopyranosyl)-1 ,3-dithio-beta-D-glucopyranose

Thiourea (5.56 g, 73.4 mmol, 5 equiv.) was added into a solution of the 2,4,6-tri-0-acetyl-3-S- (2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl)-3-thio-alpha-D-glucopyranosyl bromide (10.5 g, 14.6 mmol) from Step d. in dry acetone (1 00 mL). The reaction mixture was refluxed with TLC monitoring. After 10 minutes, the reaction mixture became a clear solution. After 2hrs, another 5 equiv. of thiourea was added and the mixture was refluxed for another 2 hours. After completion of conversion of starting material (checked by TLC), the reaction mixture was cooled to room temperature. Acetone was removed by reduced pressure. To the residue solid was added Chloroform and water (1 :1 , 300 mL) and stirred at 85 °C for 1 hour with condenser. The mixture was cooled to room temperature and extracted with DCM (3 x 200 mL). Dried and remove solvent. Crystallization in EtOH gave the title compound as a white solid (9.01 g, 92%). H NMR (300 MHz, Chloroform-d): 5.17 (dd, 1 H), 4.95-5.12 (m, 2H), 4.85-4.98 (m, 2H), 4.67 (d, 1 H), 4.43 (dd, 1 H), 4.27 (ddd, 1 H), 4.18 (m, 1 H), 4.06-4.16 (2H), 3.64-3.76 (m, 2H), 2.96 (dd, 1 H), 2.35 (d, 1 H, SH), 2.16 (s, 3H), 2.09 (s, 3H), 2.08 (s, 3H), 2.06 (s, 3H), 2.02 (s, 3H), 2.01 (s, 3H), 1 .00 (s, 3H). 13C NMR (75 MHz, Chloroform-d): 170.6 (CO), 170.5 (CO), 170.2 (CO), 169.4 (CO), 169.3 (CO), 169.1 (2CO), 84.1 (CH), 80.5 (CH), 78.5 (CH), 75.7 (CH), 75.4 (CH), 73.6 (CH), 70.1 (CH), 68.2 (CH), 66.5 (CH), 52.5 (CH), 61 .9 (CH), 52.0 (CH), 21 .0 (CH₃), 20.8 (CH₃), 20.7 (CH₃), 20.5 (3 CH₃), 20.4 (CH₃).

Step f. Synthesis of Thio-linked Trisaccharide: 2,4,6-tri-0-acetyl-3-S-(2,3,4,6-tetra-0-acetyl-beta-D- glucopyranosyl)-1 ,3-dithio-beta-D-glucopyranose

Sodium hydride (0.808 g, 29.2 mmol) was added to a solution of 2,4,6-tri-0-acetyl-3-S-(2,3,4,6- tetra-0-acetyl-beta-D-glucopyranosyl)-1 ,3-dithio-beta-D-glucopyranose (9.01 g, 13.47 mmol) in dry THF (450 mL in a 2000-mL RB Flask) at 0 °C. The suspension was stirred under nitrogen until hydrogen formation had ceased. To this solution, 1 ,7,10-trioxa-4,13-diazacyclopentadecane (Kryptofix 21 , 4,10- diaza-16-crown-5-ether, 0.535 g, 2.43 mmol) and a solution of (3aR,5R,6R,6aR)-5-((R)-2,2-dimethyl-1 ,3- dioxolan-4-yl)-2,2-dimethyltetrahydro-2H-furo[2,3-d][1 ,3]dioxol-6-yl trifluoromethanesulfonate (5.551 g, 14.1 5 mmol) in THF (200 mL) was added via cannula transfer and the mixture was stirred for 2 hours at 0 °C under Nitrogen, then at room temperature for overnight. The reaction mixture was concentrated under reduced pressure. A solution of the residue was dissolved in DCM (400 mL), washed with water (200 mL), dried (Na2S04), and concentrated. Added 40 mL of EtOH to the crude product to dissolve the foam followed by swirling the flask. The white precipitate was formed. Filtration and wash with 30% EtOAc/Hex and drying gave the title compound 6.55 g (53.4%). Check TLC of the mother liquor and some of the product remained in the mother liquor which was purified with normal phase flash column. ¹ H NMR (300 MHz, Chloroform-d): δ 5.83 (d, J = 3.5 Hz, 1 H), 5.18 (dd, J = 9.3 Hz, 1 H), 5.07 (dd, J = 10.2 Hz, 1 H), 5.06 (dd, J = 9.6 Hz, 1 H), 4.88 (ddd, J = 9.7, 9.7, 9.9 Hz, 1 H), 4.83 (d, J = 3.6 Hz, 1 H), 4.63 (dd, J = 14.2, 10.0 Hz, 2H), 4.37 - 4.21 (m, 3H), 4.21 - 4.05 (m, 5H), 4.04 - 3.96 (m, 1 H), 3.76 - 3.61 (m, 2H), 3.54 (d, J = 3.7 Hz, 1 H), 3.00 (dd, J = 10.8 Hz, 1 H), 2.15 (s, 3H), 2.10 (s, 3H), 2.08 (s, 3H), 2.08 (s, 3H), 2.02 (s, 3H), 2.01 (s, 3H), 1 .99 (s, 3H), 1 .51 (s, 3H), 1 .42 (s, 3H), 1 .34 (s, 3H), 1 .32 (s, 3H). ¹³C NMR (75 MHz, Chloroform-d): δ 170.53 (e), 170.48(e), 170.16(e), 169.42(e), 1 69.26(e), 169.09(e), 168.47(e), 1 1 1 .92(e), 109.45(e), 104.82(o), 86.04(o), 84.52(o), 84.31 (o), 80.1 1 (o), 78.43(o), 75.59(o), 73.72(o), 73.62(o), 71 .74(0), 70.02(0), 68.15(o), 67.31 (e), 66.62(o), 62.51 (e), 61 .92(e), 52.18(o), 50.07(o), 26.87(o, CH₃), 26.59(0, CH₃), 26.32(0, CH₃), 25.33(o, CH₃), 20.83(o, CH₃), 20.73(o, CH₃), 20.68(o, CH₃), 20.53(o, 3CH₃), 20.39(0, CH₃).

Step g. Synthesis of Peracetylated (1->3)-Thiotrisaccharide: 2,3,4,6-tetra-O-acetyl-beta-D- glucopyranosyl-(1->3)-2,4,6-tri-0-acetyl-3-thio-beta-D-glucopyranosyl-(1->3)-1 ,2,4,6-tetra-0-acetyl- 3-thio-D-glucopyranose

A solution of 2,4,6-tri-0-acetyl-3-S-(2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl)-1 ,3-dithio-beta- D-glucopyranose 6.55 g, 7.19 mmol) in 60% aq. AcOH/TFA (10:0.5, 105 mL) was heated for 5 hours at 70 °C, then concentrated under reduced pressure and the residue dissolved in water and freeze-dried. Conventional acetylation of the resulting solid (1 :1 Ac20/pyridine, 1 00 mL) yielded the desired product. Rotovaped to remove most of Ac20/Py, diluted with DCM (400 mL) and washed with 0.5N HCI 200 mL. 200 mL of DCM extracted the aqueous layer once. Combined DCM extract and dried with Na2S04. DCM was removed by reduced pressure. Normal phase silica column purification (0-70% EtOAc/Hex) gave the title compound (6.51 g, 90.6%) as a white solid. Step h. Synthesis of (1->3)-Thiotrisaccharide Bromide: 2,3,4,6-tetra-O-acetyl-beta-D- glucopyranosyl-(1->3)-2,4,6-tri-0-acetyl-3-thio-beta-D-glucopyranosyl-(1->3)-1 ,2,4,6-tetra-0-acetyl- 3-thio-D-glucopyranose

To a solution of the 2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl-(1 ->3)-2,4,6-tri-0-acetyl-3-thio- beta-D-glucopyranosyl-(1 ->3)-1 ,2,4,6-tetra-0-acetyl-3-thio-D-glucopyranose (6.51 g, 6.52 mmol) in dry CH2CI2 (70 mL) at 0 °C was added dropwise commercial 33% HBr in AcOH (14.2 mL, 12 equiv.). After the mixture was stirred for 3 hours at ice-bath, TLC (1 :1 EtOAc-hexane) showed complete conversion of the starting material into a single product. DCM was added (200mL), and the solution was washed with cold water (100 mL). DCM was used to extract on the aqueous layer. Combined DCM layers were washed with water (1 00 mL). Dried and removed solvent. Ethanol (Hot, 50 mL) dissolved the residue to precipitate the title compound (4.43 g, 67%). ¹ H NMR (300 MHz, chloroform-d): 6.65 (d, J = 3.63, 1 H), δ 5.18 (dd, J = 9.3 Hz, 1 H), 5.02 - 5.13 (m, 2H), 4.80 - 5.01 (m, 4H), 4.72 (d, J = 9.7, 1 H), 4.65 (d, J = 1 0.2, 1 H), 4.07 - 4.35 (m, 7H), 3.67 - 3.78 (m, 2H), δ 3.32 (dd, J = 1 1 .1 Hz, 1 H), 3.00 (dd, J = 10.5 Hz, 1 H), 2.16 (s, 3H), 2.14 (s, 3H), 2.12 (s, 6H), 2.1 1 (s, 3H), 2.10 (s, 3H), 2.08 (s, 3H), 2.04 (s, 3H), 2.02 (s, 3H), 2.00 (s, 3H). ¹³C NMR (75 MHz, Chloroform-d): δ 170.55 (e), 170.53 (e), 170.50 (e), 1 70.21 (e), 169.43 (e), 169.40(e), 169.38 (e), 169.30 (e), 169.10 (e), 168.53 (e), 89.00 (0), 84.23 (0), 77.82 (0), 75.58 (0), 73.61 (0), 73.61 (0), 73.00 (0), 71 .83 (0, 2C), 71 .48 (0), 69.96 (0) , 68.01 (0), 66.41 (0), 64.65 (0), 62.32 (e), 61 .86 (e), 61 .51 (e), 52.17 (0), 46.83 (0), 20.78 (0), 20.74 (0, 2C), 20.67 (0), 20.64 (0), 20.60 (0, 3C), 20.54 (0), 20.44 (0). Step i. Synthesis of (1->3)-Thiotrisaccharide Thiol: 2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl-(1- >3)-2,4,6-tri-0-acetyl-3-thio-beta-D-glucopyranosyl-(1->3)-2,4,6-tri-0-acetyl-1 ,3-dithio-beta-D- glucopyranose

A similar procedure was used as that in Example 3, Step e. Crystallization from EtOH gave 3.50 g of the title compound as a white solid. Column purification of mother liquor gave a white solid (0.55 g). The yield was 95.9%. Ή NMR (300 MHz, Chloroform-d): δ 5.16 (dd, J = 9.3 Hz, 1 H), 4.99 -5.12 (m, 2H), 4.79 - 4.80 (m, 4H), 4.64 (d, J = 9.9, 1 H), 4.61 (d, J = 4.0, 1 H), 4.41 (t, J = 4.5 Hz, 1 H), 4.30 - 4.04 (m, 6H), 3.60 - 3.75 (m, 3H), 2.85 - 3.00 (m, 2H), 2.38 (d, J = 9.9 Hz, 1 H), 2.14 (s, 3H), 2.10 (s, 3H), 2.08 (s, 9H), 2.07 (s, 6H), 2.01 (s, 3H), 1 .99 (s, 3H), 1 .98 (s, 3H). ¹³C NMR (75 MHz, Chloroform-d): δ 170.68 (e), 170.64 (e), 170.55 (e), 170.17 (e), 169.50 (e), 169.39 (e), 169.29 (e), 169.1 0 (e), 1 68.95 (e), 1 68.56 (e), 84.87 (0), 84.10 (0), 80.73 (0), 78.45 (0), 77.84 (0), 75.55 (0), 74.99 (0), 73.60 (0), 71 .84 (0), 69.96 (0), 68.04 (0), 66.65 (0), 66.14 (0), 62.49 (e, 2C), 61 .84 (e), 52.12 (0), 51 .84 (0), 20.98 (0), 20.88 (0), 20.77 (0), 20.74 (0), 20.65 (0), 20.59 (0), 20.55 (0, 3C), 20.41 (0). Step j. Synthesis of Peracetylated (1->3)-Thiotrisaccharide ethyleneamine: 2-aminoethyl 2,3,4,6- tetra-0-acetyl-beta-D-glucopyranosyl-(1->3)-2,4,6-tri-0-acetyl-3-thio-beta-D-glucopyranosyl-(1->3)- 2,4,6-tri-0-acetyl-1 ,3-dithio-beta-D-glucopyranoside (INT-3)

Part 1 (Mitsunobu Reaction). Trimethylphosphine (TMP, 1 .0M in THF, 2.1 ml_, 2.06 mmol) was added to a solution of 1 ,1 '-(azodicarbonyl)dipiperidine (ADDP, 0.519 g, 2.06 mmol) in 25 mL of THF at 0°C and stirred for 30 min. N-Cbz-ethanolamine (0.201 g, 1 .03 mmol) and 2,3,4,6-tetra-O-acetyl-beta-D- glucopyranosyl-(1 ->3)-2,4,6-tri-0-acetyl-3-thio-beta-D-glucopyranosyl-(1 ->3)-2,4,6-tri-0-acetyl-1 ,3-dithio- beta-D-glucopyranose (0.54 mmol in 5 mL of THF) were added sequentially to the solution, with further stirring at room temperature for 2 hours. Any precipitate was then filtered off and the solution evaporated to dryness. Normal phase silica gel column purification (0-70% EtOAc/Hex) gave the desired product as a white foam (0.946 g, 80%). ¹³C NMR (75 MHz, Chloroform-d): δ 170.70 (e), 1 70.59 (e), 1 70.57 (e), 170.20 (e), 169.46 (e), 169.36 (e), 169.30 (e), 169.12 (e), 168.66 (e), 168.57 (e), 1 56.27 (e), 136.44 (e), 128.54 (o, 2C), 128.19 (o, 3C), 86.05 (o), 85.16 (o), 84.10 (o), 78.1 1 (o), 77.75 (o), 75.56 (o), 73.60 (o), 71 .73 (o), 71 .23 (o), 69.95 (o), 68.04 (o), 66.70 (e), 66.59 (o), 66.29 (o), 62.46 (e), 61 .85 (e), 52.18 (o), 51 .81 (o), 41 .13 (e), 31 .35 (e), 20.94 (o), 20.85 (o), 20.77 (o), 20.67 (o, 2C), 20.61 (o), 20.58 (o, 2C), 20.55 (o), 20.44 (o).

Part 2 (Hydrogenolysis). The compound from Example 3, Step j, Part 1 (0.500 g, 0.435 mmol) and 30% Pd/C (0.463 g, 1 .30 mmol) were in the flask with 20 mL of EtOH and 5 mL of CHCI₃. Under H₂, the reaction mixture was stirred overnight. Check TLC and mass. Celite filtration and wash with EtOH gave the title compound 0.44 g (100%). Mass showed a strong detectable positive charge signal at tr=3 min with 8 min (5~95%)/highMW method (found M+H⁺: 101 6.2).

INT-3 (1 eq) is dissolved in methanol and excess NaOMe (10 eq) is added. The reaction is stirred until LCMS shows substantial formation of the desired product (INT-4) with an exact mass of 595.14.

Example 5. Synthesis of (1->3)-Thiotrisaccharide ethyleneamine: 4-[(2-{[beta-D-glucopyranosyl-(1 >3)-3-thio-beta-D-glucopyranosyl-(1->3)-3-thio-beta-D-glucopyranosyl]thio}ethyl)amino]-4- oxobutanoic acid (INT-5)

Step a. Succinic anhydride (0.0879 g, 0.869 mmol) was added to INT-3 (0.4417 g, 0.435 mmol) in DCM/Py (20 mL). The reaction mixture was stirred at room temperature overnight. LC/MS 8min Positive/negative showed the desired peak mass at 4.3 min on negative ionization. The solvent was removed and the residue was loaded onto an Isco Gold 24g column and eluted with 4-6%MeOH/DCM to give 0.27 g of the desired product. ¹³C NMR (75 MHz, Chloroform-d) δ 172.68, 1 70.94, 170.88, 170.65, 170.23, 169.62, 169.44, 169.35, 169.16, 168.99, 168.64, 85.52, 85.30, 84.08, 78.1 0, 77.57, 75.52, 73.60, 71 .75, 71 .32, 69.95, 68.03, 66.60, 66.41 , 62.56, 61 .86, 52.1 6, 51 .87, 50.68, 39.40, 30.89, 30.22, 29.68, 20.98, 20.86, 20.78, 20.66, 20.63, 20.58, 20.43. Step b. NaOMe in MeOH (25%, 1 .42 mL, 25 equiv.) was added into a 50-mL RB Flask which contained the product of Step a (0.270 g, 0.242 mmol) in 5 mL of MeOH. The mixture was stirred at room temperature for 60 hours. It was then acidified with 1 N aq. HCI and purified with reversed phase C18 column (0-3% CAN/water) to give 0.168 g (99.8%) of the title compound (INT-5). LC/MS 8 min pos/neg method showed negative mass at the very beginning (M-H, 694.2). 13C NMR (75 MHz, water-d): δ 181 .01 (e), 175.85 (e), 86.73 (o), 85.96 (o), 84.51 (o), 81 .72 (o), 81 .58 (o), 79.67 (o), 77.08 (o), 72.57 (o), 72.24 (o), 71 .81 (o), 69.27 (o), 66.88 (o, 2C), 61 .02 (e), 60.92 (e), 60.59 (e), 56.01 (o), 55.76 (o), 39.36 (e), 32.92 (e), 32.25 (e), 29.10 (e).

Step a. Synthesis of 1 ,2,3,4-tetra-0-acetyl-6-deoxy-6-iodo-beta-D-glucopyranose

Iodine (9.42 g, 37.1 mmol) was added at room temperature to a mixture of 1 ,2,3,4-tetra-O-acetyl- beta-D-glucose (9.93 g, 28.55 mmol) and chlorodiphenylphosphine (CIDPP, 6.82 mL, 37.1 mmol) and imidazole (4.28 g, 62.8 mmol) in toluene (-100 mL). After 5 minutes of stirring the reaction mixture started to produce heat. The temperature rose to about 50 °C. After 30 minutes the reaction mixture was heated at 50 °C for 6 hours when TLC indicated that the reaction was complete. Workup: The reaction mixture was cooled to room temperature and poured into an equal volume of saturated Na2C03 in a separatory funnel. The funnel was shaken for 5 minutes while iodine was added portion wise until the toluene phase remained iodine-colored. The aqueous layer was separated and the organic layer was washed with saturated aqueous Na2S203 to remove the excess iodine and then washed with water and brine. Dried and removed the solvent. Purified with 0-40% EA/Hex to give 8.02 g of iodide 2 as a white solid (61 %). ¹ H NMR (300 MHz, Chloroform-d) δ 5.77 (d, J = 8.2 Hz, 1 H), 5.28 (dd, J = 9.4 Hz, 1 H), 5.15 (dd, J = 9.4, 8.2 Hz, 1 H), 5.01 (dd, J = 9.4 Hz, 1 H), 3.59 (ddd, J = 9.54 6.4, 3.0 Hz, 1 H), 3.35 (dd, J = 1 1 .3, 3.0 Hz, 1 H), 3.1 8 (dd, J = 1 1 .3, 6.4 Hz, 1 H), 2.15 (s, 3H), 2.08 (s, 3H), 2.05 (s, 3H), 2.04 (s, 3H).

Step b. Synthesis of (1->6)-Thiodisaccharide: 1 ,2,3,4-tetra-0-acetyl-6-S-(2,3,4,6-tetra-0-acetyl- beta-D-g lucopyranosyl)-6-th io-beta-D-g lucopyranose

Sodium hydride (0.576 g, 14.4 mmol) was added to a solution of 2,3,4,6-tetra-0-acetyl-l-thio-p-D- glucopyranose (5.01 g, 13.75 mmol) in dry THF (130 mL) at 0 °C. The suspension was stirred under nitrogen until hydrogen formation had ceased. The resulting solution was then concentrated under reduced pressure, and the residue was dissolved in 60 mL of DMF. To this solution, 1 ,2,3,4-tetra-O- acetyl-6-deoxy-6-iodo-beta-D-glucopyranose (1 eq in 30 mL of DMF) was added. The mixture was stirred for 3 hours at room temperature under nitrogen, then concentrated under reduced pressure. A solution of the residue in DCM (250 mL) was washed with water (50 mL x 2), dried (Na2S04), and concentrated. Crystallization of the crude product was accomplished from EtOH. After drying, 7.23 g of the title compound as a white powder was obtained (79%). ¹ HNMR showed that there was no alpha isomer. Mother liquor was purified by silica column. After removal of solvents, the residue was subjected to EtOH precipitation and 0.37 g of a white crystalline solid was obtained (4.1 %). Both materials showed the correct ¹ HNMR and ¹³CNMR, identical to the reference.

Step c. Synthesis of (1->6)-Thiodisaccharide Bromide: 2,3,4-tri-0-acetyl-6-S-(2,3,4,6-tetra-0- acetyl-beta-D-glucopyranosyl)-6-thio-alpha-D-glucopyranosyl bromide

To a solution of 1 ,2,3,4-tetra-0-acetyl-6-S-(2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl)-6-thio- beta-D-glucopyranose (7.600 g, 10.94 mmol) in dry DCM (120 mL) at 0 °C was added dropwise commercial 33% HBr in AcOH (23.8 mL, 12 equiv.). After the mixture was stirred for 3 hours in an ice- bath, TLC (1 :1 EtOAc-hexane) showed complete conversion of the starting material into a single product. DCM was added (240 mL), and the solution was washed with cold water (2 x 100 mL). After removal of the solvents, the residue was subjected to EtOH precipitation and 6.87 g of the title compound (as a white crystalline solid) was obtained (87.8%). Both showed the correct ¹ HNMR and ¹³CNMR, identical to the reference. Step d. Synthesis of (1->6)-Thiodisaccharide Thiol: 2,3,4-tri-0-acetyl-6-S-(2,3,4,6-tetra-0-acetyl- beta-D-glucopyranosyl)-1 ,6-dithio-beta-D-glucopyranose

A similar procedure was used as that in Example 3, Step e. Crystallization from EtOH gave 3.74 g of the title compound (white solid) from 4.27 g of 2,3,4-tri-0-acetyl-6-S-(2,3,4,6-tetra-0-acetyl-beta-D- glucopyranosyl)-6-thio-alpha-D-glucopyranosyl bromide (94% yield). ¹ HNMR and ¹³CNMR were identical to the reference (Carbohydrate Research, 281 (1996) 99-1 18).

Step e. Synthesis of (1->6)-Thiotrisaccharide: 2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl-(1->6)- 2,3,4-tri-0-acetyl-6-thio-beta-D-glucopyranosyl-(1->6)-1 ,2,3,4-tetra-0-acetyl-6-thio-beta-D- glucopyranose

Reaction and workup procedure are analogous to those described above in Step b. Purification: normal phase silica gel column 0-90% purify the crude residue. The collection was clean by TLC but contained NMP. After removing EA/Hex, added water and lyophilized twice to remove NMP. ¹ HNMR is identical to the reference (Carbohydrate Research, 281 (1996) 99-1 18). 4.62 g of the title compound was obtained (73% based on bromide).

Step f. Synthesis of (1->6)-Thiotrisaccharide Bromide: 2,3,4,6-tetra-O-acetyl-beta-D- glucopyranosyl-(1->6)-2,3,4-tri-0-acetyl-6-thio-beta-D-glucopyranosyl-(1->6)-2,3,4-tri-0-acetyl-6- thio-alpha-D-glucopyranosyl bromide

An analogous procedure to that used in Step c of this Example was used. 4.26 g of the title compound was obtained as a white solid (82%). ¹³C NMR (75 MHz, CDC ): δ 170.69 (e), 170.12 (e, 2C), 169.83 (e), 169.76 (e), 169.70 (e), 169.64 (e), 169.49 (e), 169.44 (e), 169.37 (e), 86.17 (o), 83.48 (o), 83.1 8 (o), 78.26 (o), 76.00 (o), 74.33 (o), 73.72 (o), 73.56 (o), 71 .52 (o), 70.67 (o), 70.43 (o), 70.09 (o), 70.00 (o), 69.87 (o), 68.20 (o), 62.07 (e), 31 .07 (e), 30.17 (e), 20.84 (o, CH₃), 20.76 (o, 3 CH₃), 20.70 (o, CH₃), 20.66 (o, CH3), 20.65 (o, CH₃), 20.61 (o, 3 CH₃).

Step g. Synthesis of (1->6)-Thiotrisaccharide Thiol: 2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl-

(1->6)-2,3,4-tri-0-acetyl-6-thio-beta-D-glucopyranosyl-(1->6)-2,3,4-tri-0-acetyl-1 ,6-dithio-beta-D- glucopyranose

An analogous procedure as that used in Example 3, Step e was used. 3.20 g of the title compound was obtained as a white solid (75%). ¹ H NMR (300 MHz, Chloroform-d): δ 5.10 -5.29 (m, 3H), 4.91 - 5.07 (m, 5H), 4.74 (d, J = 10.1 , 1 H), 4.68 (d, J = 1 0.4, 1 H), 4.62 (dd, J = 9.8 Hz, 1 H), 4.30 (dd, J = 4.9, 12.4, 1 H), 4.18 (dd, J = 1 .6, 1 1 .0, 1 H), 3.74 - 3.84 (m, 2H), 3.65 - 3.73 (m, 2H), 2.79 - 2.90 (m, 2H), 2.42 (d, J = 9.8 Hz, 1 H), 2.15 (s, 3H), 2.1 1 (s, 3H), 2.1 0 (s, 3H), 2.09 (s, 6H), 2.08 (s, 3H), 2.05 (s, 3H), 2.03 (s, 6H), 2.02 (s, 3H). ¹³C NMR (75 MHz, CDCI₃) δ 170.69 (e), 170.17 (e), 170.13 (e), 170.1 1 (e), 169.65 (e), 169.62 (e, 2C), 169.42 (e), 169.40 (e), 169.36 (e), 83.44 (o), 83.12 (o), 78.92 (o), 78.54 (o), 77.93 (o), 76.10 (o), 73.73 (o), 73.65 (o, 2C), 73.40 (o), 71 .52 (o), 71 .40 (o), 70.44 (o), 69.74 (o), 68.1 7 (o), 62.04 (e), 31 .23 (e), 31 .05 (e), 20.85 (o), 20.79 (o, 2C), 20.76 (o, 2C), 20.71 (o), 20.63 (o, 4C). Step h. Synthesis of Peracetylated (1->6)-Trisaccharide ethyleneamine: 2-aminoethyl 2,3,4,6-tetra- 0-acetyl-beta-D-glucopyranosyl-(1->6)-2,3,4-tri-0-acetyl-6-thio-beta-D-glucopyranosyl-(1->6)-2,3,4- tri-0-acetyl-1 ,6-dithio-beta-D-glucopyranoside

Part 1 (Mitsunobu Reaction). The reaction was carried out in a similar manner as Example 3, Step j, Part 1 except the purification method was different. Reversed phase C18 column (10-80%

ACN/water with 0.1 %TFA) was used. 1 .02 g of the desired (1 ->6) product was obtained (87%) from 2,3,4,6-tetra-0-acetyl-beta-D-glucopyranosyl-(1 ->6)-2,3,4-tri-0-acetyl-6-thio-beta-D-glucopyranosyl-(1 - >6)-2,3,4-tri-0-acetyl-1 ,6-dithio-beta-D-glucopyranose (1 .00 g 1 .03 mmol). ¹³C NMR (75 MHz, CDC ) δ 170.66 (e), 170.18 (e), 170.13(e), 1 70.06 (e), 1 69.67(e), 169.61 (e), 169.44 (e, 2C), 169.40 (e), 169.34 (e), 156.33 (e), 136.57 (e), 136.40 (e), 128.51 (o), 128.10 (o, 2C), 83.76 (o, 2C), 83.53 (o), 77.57 (o), 76.20 (o), 73.65 (o), 73.49 (o, 2C), 73.45 (o), 71 .49 (o), 71 .45 (o), 70.1 6 (o), 70.13 (o), 69.61 (o), 68.1 5 (o), 66.81 (e), 66.62 (e), 62.17 (e), 62.1 0 (e), 43.47 (e), 41 .05 (e), 20.83 (o, CH₃), 20.78 (o, CH₃), 20.73 (o, CH₃), 20.69 (o, 2 CH₃), 20.60 (o, 5 CH₃).

Part 2 (Hydrogenolysis). The reaction was carried out in a similar manner as Example 3, Step j, Part 2 to obtain INT-6. Ion found by LCMS (M+H⁺: 101 6.1 ).

Exampl -7

In a manner similar to that described for the synthesis of INT-4 but using INT-6 as the starting material, INT-7 is obtained having an exact mass of 595.14.

Example 8. Synthesis of (1->6)-Thiotrisaccharide ethyleneamine: 4-[(2-{[beta-D-glucopyranosyl- (1->6)-6-thio-beta-D-glucopyranosyl-(1->6)-6-thio-beta-D-glucopyranosyl]thio}ethyl)amino]-4- oxobutanoic acid (INT-8)

Step a. An analogous procedure was used as that for the preparation of the compound from Example 5, Step a. The desired compound (0.480 g) was obtained in 99% yield. ¹³C NMR (75 MHz, CDCI₃): δ 172.65 (e), 170.75 (e), 170.22 (e), 1 70.1 7 (e), 170.09 (e), 169.78 (e), 169.75 (e, 3C), 169.57 (e), 169.49 (e), 169.46 (e), 83.85 (o), 83.63 (o), 83.55 (o), 77.49 (o), 77.43 (o), 76.24 (o), 73.65 (o), 73.43 (o), 73.38 (o), 71 .58 (o), 71 .50 (o), 70.25 (o, 2C), 69.59 (o), 68.16 (o), 62.14 (e), 39.69 (e), 32.29 (e), 31 .29 (e), 30.78 (e), 30.38 (e, 2C), 20.86 (o,CH₃), 20.79 (o,CH₃), 20.75 (o, CH₃), 20.72 (o, 3 CH₃), 20.62 (o, 4 CH₃). Step b. An analogous procedure was used as that for the preparation of the compound from Example 5, Step b. INT-8 (0.275 g, 99% yield) was obtained. Negative ion was found (M-H-: 694.0). ¹³C NMR (75 MHz, D₂0): δ 180.94 (e), 175.76 (e), 87.07 (0) , 86.04 (0) , 85.34 (0) , 79.74 (0) , 79.23 (0) , 79.20 (0) , 77.14 (0) , 76.87 (0, 2C), 72.70 (0) , 72.46 (0) , 72.41 (0, 2C), 72.33 (0) , 69.37 (0) , 60.77 (e), 39.43 (e), 33.07 (e), 32.94 (e), 32.30 (e), 32.01 (e), 29.51 (e).

Example 9. Synthesis of INT-9

reflux 85°C

Step a. Synthesis of benzyl [2-(2-hydroxyethoxy)ethyl]carbamate

Triethylamine (1 .31 mL, 9.32 mmol) was added to a cold (0 °C) solution of 2-(2- aminoethoxy)ethan-1 -ol (CAS# 929-06-6, 1 .00 g, 9.32 mmol) in anhydrous THF (100 mL). To the mixture was then added slowly Cbz chloride (1 .50 mL, 10.3 mmol). After stirring at room temperature for 1 hour, the reaction was diluted with ethyl acetate (250 mL) and the organic layer was washed with a mixture of saturated sodium bicarbonate and brine (50 mL + 50 mL), dried (Na2S04) and concentrated under reduced pressure to provide the crude product. The crude residue was purified by normal phase silica flash column (0-10% EA/Hex) to give the desired product (14.1 5 g, 89%) as a colorless oil. ¹ HNMR (300MHz, Chloroform-d): δ 7.45 - 7.29 (m, 5H), 5.1 8 (bs, 1 H, NH), 5.13 (m, 2H), 3.75 (td, J = 5.6, 3.9 Hz, 2H), 3.59 (m, 4H), 3.43 (td, J = 5.6, 3.9 Hz, 2H), 2.06 (t, J = 6.0 Hz, 1 H, OH). Step b. Synthesis of Per-acetyl Rhamnose Bromide: 2,3,4-tri-0-acetyl-6-deoxy-beta-L- mannopyranosyl bromide

To a solution of L-rhamnose (69.0 g, 0.3799 mol, 1 .0 eq) and DMAP (1 .40 g, 0.02x) in pyridine (280mL, 4V/M) was added acetic anhydride (232.0 g, 2.273 mol, 6.0 equiv) dropwise with cooling by an ice-water bath (control the temperature below 20°C). The mixture was stirred at ambient temperature (12 hrs), and volatiles were removed under reduced pressure (toluene azeotrope). The crude product was dissolved in EtOAc (800 mL), washed with hydrochloric acid (1 N, 800 mL^*2), sat. NaHCO3 (800 mL), brine (800 mL), and dried with sodium sulfate (100g). Volatiles were removed under reduced pressure, and crude acetate (131 .0g, yield: 100%) was used directly for the next step. To a solution of the crude peracetylated rhamnose (6.34 g, 15.3 mmol) in dry DCM (150 mL) at 0 °C was added dropwise commercial 33% HBr in AcOH (33.2 mL, 12 equiv.). After the mixture was stirred for 3 hours at ice-bath, TLC (1 :1 EtOAc-hexane) showed complete conversion of the mixed SM into a single product. DCM was added (240mL), and the solution was washed with cold water (2 x 100 mL) and dried with Na2S04. DCM was removed by reduced pressure. The residue was purified using silica column (0-50% EA/Hex) to give a white solid (Rf = 0.5 at 30%EA/Hex) in 84% yield.

Step c. Synthesis of Peracetylated Rhamnosyl Thiol: 2,3,4-tri-0-acetyl-6-deoxy-1-thio-alpha-L- mannopyranose

Thiourea (2.82 g, 37.0 mmol, 5 equiv.) was added into a solution of the above 2,3,4-tri-O-acetyl-

6-deoxy-beta-L-mannopyranosyl bromide in dry acetone (40mL). The reaction mixture was refluxed with TLC monitoring. After 10 minutes, the reaction mixture became a clear solution. After 2 hrs, another 5 equiv. of thiourea was added and the mixture was refluxed for another 2 hours. After completion of conversion of starting material (checked by TLC), the reaction mixture was cooled to room temperature. Acetone was removed under reduced pressure. To the solid residue was added chloroform and water (1 :1 , 300 mL) and stirred at 85 °C for 1 hour with condenser. The mixture was cooled to room

temperature and extracted with DCM (3 x 200 mL), the DCM layer dried, filtered and the solvent removed. The residue was purified by silica gel chromatography (0-50% EA/Hex) to give a white solid (Rf = 0.5 at 30%EA/Hex) in over 80% yield. ¹ H NMR (300 MHz, Chloroform-d): δ 5.49 (dd, J = 6.9, 1 .3 Hz, 1 H), 5.33 (s, 1 H), 5.38 - 5.26 (m, 1 H), 5.1 0 (t, J = 9.4 Hz, 1 H), 4.30 - 4.06 (m, 1 H), 2.26 (d, J = 7.0 Hz, 1 H), 2.1 7 (s, 3H), 2.08 (s, 3H), 2.01 (s, 3H), 1 .52 (d, J = 6.2 Hz, 3H). 13C NMR (75 MHz, Chloroform-d): 170.0 (CO), 170.0 (CO), 169.3 (CO), 84.1 (CH), 76.7 (CH), 72.3 (CH), 71 .0 (CH), 69.5 (CH), 67.6 (CH), 20.9 (CH₃), 20.8 (CH₃), 20.7 (CH₃), 17.3 (CH₃). Step d. Synthesis of Rhamnose-PEG1-NHCbz: benzyl (2-{2-[(2,3,4-tri-0-acetyl-6-deoxy-alpha-L- mannopyranosyl)thio]ethoxy}ethyl)carbamate

Trimethylphosphine (TMP, 1 .0M in THF, 1 .3 mL, 1 .3 mmol) was added to a solution of 1 ,1 '- (azodicarbonyl)dipiperidine (ADDP, 0.329 g, 1 .3 mmol) in 2 mL of THF at 0 °C and stirred for 30 min. The Cbz-2-(2-hydroxyethoxy)ethyl alcohol (0.164 g, 0.686 mmol) from Step a and the 2,3,4-tri-0-acetyl-6- deoxy-1 -thio-alpha-L-mannopyranose from Step b (in 2 mL of THF) were added sequentially to the solution, with further stirring at room temperature for 2 hours. Any precipitate was then filtered off and the solution evaporated to dryness. Normal phase silica gel column purification (0-70% EtOAc/Hex) gave the desired product as a white foam (0.946 g, 80%). ¹ H NMR (300 MHz, Chloroform-d): δ 7.42 - 7.28 (m, 5H), 5.40 - 5.25 (m, 2H, NH, CH), 5.28 (d, J = 1 .5 Hz, 1 H), 5.22 (dd, J = 10.0, 3.3 Hz, 1 H), 5.17 - 5.05 (m, 3H), 4.20 (m, 1 H), 3.64 (m, 2H), 3.53 (m, 2H), 3.38 (m, 2H), 2.84 (m, 1 H), 2.72 (m, 1 H), 2.13 (s, 3H), 2.05 (s, 3H), 1 .99 (s, 3H), 1 .23 (d, J = 6.2 Hz, 3H). ¹³C NMR (75 MHz, CDCI3) δ 170.19 (e), 169.99 (e), 169.88 (e), 156.47 (e), 136.57 (e), 128.50 (o, 2C), 128.07 (o, 3C), 82.64 (o), 77.52 (CDCI3), 77.09 (CDCI3), 76.67 (CDCI3), 71 .52 (o), 71 .1 5 (o), 70.55 (e), 69.94 (e), 69.24 (o), 67.1 9 (o), 66.64 (e), 40.87 (e), 30.72 (e), 20.95 (o), 20.82 (o), 20.70 (o), 1 7.37 (o). Step e. Synthesis of Rhamnose-PEG1-NH Succinic Anhydride Adduct: 4-oxo-4-[(2-{2-[(2,3,4-tri-0- acetyl-6-deoxy-alpha-L-mannopyranosyl)thio]ethoxy}ethyl)amino]butanoic acid

Part 1 (Hydrogenolysis): Benzyl (2-{2-[(2,3,4-tri-0-acetyl-6-deoxy-alpha-L- mannopyranosyl)thio]ethoxy}ethyl)carbamate (0.200 g, 0.379 mmol) and 30% Pd/C (0.134 g, 0.379 mmol) were loaded in the flask with 8 mL of EtOAc and 2 ml_ of CHCI₃. Under H2, the reaction mixture was stirred overnight. Check TLC and mass spectrum. The material was filtered through celite and washed with EtOH to give the title compound 0.14 g. Mass spectrum showed a strong detectable positive charge signal (5-95%) (found M+H⁺: 394.2 and M-Boc+H⁺: 293.2). Part 2 (Succinic anhydride adduct): To the material from Part 1 in DCM/Py (10 mL) was added succinic anhydride (0.540 g, 0.534 mmol, 1 .5 equiv.). The reaction mixture was stirred at room temperature overnight. LC/MS 8min Positive/negative showed the desired peak mass at 3.72 min on both positive (M+H⁺: 494.2) and negative ionization (492.2). The solvent was removed and the residue was loaded onto a silica gel column and eluted with 4-6% MeOH/DCM to give the title compound. ¹³C NMR (75 MHz, CDCI3) δ 175.94 (CO), 172.53 (CO), 170.42 (CO), 170.02 (2CO), 82.53 (CH), 71 .63 (CH), 71 .08 (CH), 70.51 (CH₂), 69.60 (CH₂), 69.25 (CH), 67.20 (CH), 39.38 (CH₂), 30.82 (CH₂), 30.71 (CH₂), 28.9 (CH₂), 21 .00 (CH₃), 20.80 (CH₃), 20.70 (CH₃), 17.36 (CH₃).

Step f. Synthesis of Rhamnosyl-PEG1-Succinic acid linker: 4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)thio]ethoxy}ethyl)amino]-4-oxobutanoic acid (INT-9)

NaOMe in MeOH (0.33 mL, 1 .42 mmol) was added into the 20-mL vial which contained the product of Step e (0.140 g, 0.284 mmol) in 3 mL of MeOH. Stirred at room temperature for overnight. LC/MS 5 min pos/neg method showed desired mass at 2.01 7 min (M+H⁺: 368.2 and M-H-: 366.2).

Acidified to pH~3 with AcOH and the solvent was evaporated under reduced pressure. The residue as purified by C1 8 reverse phase chromatography to give the title compound INT-9.

Example 10. Synthesis of INT-10 (Gala1-3Galp1-4GlcNAcp-OCH₂CH₂NH₂)

Step b. Synthesis of galactose coupling partner

Step c. Synthesis of protected 2-glucosamine-1-azidoethylether

Step d. Synthesis of INT-10 (Gala1-3Galp1-4GlcNAcp-OCH₂CH₂NH₂)

INT-10 (Galal -3Galp1 -4GICNACP-OCH2CH2NH2) was prepared in a manner similar to that described in WO 2014/151423 A1 (Example 1 , Scheme 5) with the modifications noted in the preceding scheme detailing the preparation. ¹ H NMR (400 MHz, D2O) : δ 5.03 (d, 3.6 Hz, 1 H), 4.41 (d, 7.6 Hz, 2H), 4.10-4.04 (m, 2H), 4.06-3.49 (m, 15H). 2.73-2.71 (m, 2H), 1 .93 (s, 3H). LCMS: m/z calcd for

C22H40N2O16: 588.24; found: 589.2 [M+H]⁺ Example 11. Degradation of polymyxin B to tri-Boc polymyxin B cycloheptapeptide (INT-11 )

polymyxin B

INT-11

Polymyxin B (100 g, 72.2 mmol) was dissolved in acetonitrile (1 000 mL, 10V) and water (500 mL, 5V) and stirred at room temperature for 10 mins. TEA (58.5 g, 8.0 eq) was added and the mixture stirred for a further 10 mins. B0C2O (94.6 g, 6.0 eq) was subsequently added in one portion and the mixture stirred for 6 hrs at 20 °C. Savinase (300 mL) was then added. The pH of the resulting mixture was adjusted to 9.0 with aq 4M sodium hydroxide solution (10 mL) and the reaction mixture stirred at 25°C. Additional savinase (100 mL, 1 V) was added after 17 hrs, and another quantity of savinase (100 mL x2, 1 VX2) was added after 26 hrs. After an overall reaction time of 80 hrs, the mixture was diluted with EA (2000 mL, 20V). After separation of the layers, the organic phase was washed with 0.1 M NaOH solution (1000 mL x2, 10V x2), then water (1000 mL, 10V). The organic layer was dried over anhydrous Na2S04, filtered and the solvent evaporated at reduced pressure. The residue was purified by silica gel chromatography eluting with 80% (EtOAc : MeOH : Η₂ΟΝΗ₃ = 40:1 0:1 ) in ethyl acetate to give the title compound INT-1 1 (49.8 g, 65.0%). LCMS: m/z (M + H)⁺ calcd for CsoHeaNnO :! 061 .61 ; found:1062.5

Example 12. Degradation of polymyxin E to tri-Boc polymyxin E cycloheptapeptide (INT-12)

INT-12

Procedure A

Colistin sulfate (5.0 g, 3.95 mmol) was dissolved in acetonitrile (50 mL) and water (25 mL) and stirred at room temperature for 10 minutes. Triethylamine (3.2 mL, 23.0 mmol) was added and the mixture stirred for a further 10 minutes. Di-tert-butyl dicarbonate (5.0 g, 23.0 mmol) was subsequently added in one portion and the mixture stirred for 16 hours. Savinase (Novozymes) (15 mL) was then added, followed by 4 M sodium hydroxide solution (0.5 mL) and the reaction mixture stirred at room temperature for 5 days. The mixture was diluted with ethyl acetate and water. After separation of the layers, the organic phase was washed with 0.1 M sodium hydroxide solution (x2), then water. The organic layer was dried over magnesium sulfate, filtered and the solvent evaporated at reduced pressure. The residue was purified by reversed phase chromatography (1 0-95% acetonitrile/di water containing 0.1 % formic acid: 25 minute gradient). The pure fractions were pooled and lyophilized to afford title compound INT-12 as a formate salt, white powder. (2.28 g, 56%). m/z 1028, [M+H]⁺

Procedu e B

Colistin sulfate (50.0 g, 39.5 mmol) was dissolved in acetonitrile (500 mL, 10V) and water (250 mL, 5V) and stirred at room temperature for 10 mins. TEA (24.2 g, 6.0eq) was added and the mixture stirred for a further 10 mins. B0C2O (52.2 g, 6.0 eq) was subsequently added in one portion and the mixture stirred for 29 hrs. LCMS showed material no being detected. Savinase (1 50 mL) was then added. The pH of the resulting mixture was adjusted to 9.0 with aq 4M sodium hydroxide solution (5 mL) and the reaction mixture stirred at 25°C. Additional savinase (50 mL, 1 V) was added after 79 hrs, and another quantity of savinase (50 mL, 1 V) was added after 103 hrs. After an overall reaction time of 162 hrs, the mixture was diluted with EA (1000 mL, 20V). After separation of the layers, the organic phase was washed with 0.1 M NaOH solution (500 mL x2, 10V x2), then water (500 mL, 10V). The organic layer was dried over anhydrous Na2S04, filtered and the solvent evaporated at reduced pressure. The residue was purified by silica gel chromatography eluting with 80% (EtOAc:MeOH:NH4OH=40:10:1 ) in ethyl acetate to give the title compound INT-12 (20.2 g, 49.3%). LCMS: m/z (M + H)⁺ calcd for 047Η₈₅ΝιιΟΐ4:1027.63; found:1028.5.

Example 13. Preparation of tetra-Boc Dab-polymyxin E cycloheptapeptide (INT-13)

Step a. Synthesis of Cbz-L-Dab-IN -12

INT-12 (0.20 g, 0.195 mmol) , Z-L-Dab(Boc)-OH-DCHA (0.124 g, 0.233 mmol), and HOBT (0.029 g, 0.214 mmol) were dissolved in DMF (2 mL) and EDC (0.041 g, 0.214 mmol) was added and the reaction was stirred for 2 hours. The mixture was applied directly to reversed phase HPLC (25-95% acetonitrile in Dl water containing 0.1 % formic acid:20 minute gradient). The pure fractions were lyophilized to afford 0.197 g of the title compound as a white solid. Yield: 74%. LC/MS (M+2H)/2 = 682.0. Step b. Synthesis of INT-13

The Cbz-L-Dab-INT-12 (0.197 g, 0.194 mmol) was stirred in methanol (15 mL) in the presence of 5% Pd/C (50 mg) under 1 atm of hydrogen gas for 45 minutes. The mixture was filtered, concentrated and applied to reversed phase HPLC (25-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were lyophilized to afford 0.140 g of the title compound as a white solid. Yield: 72%. LC/MS (M+2H)/2= 565.0 (loss of 1 Boc in LC/MS).

Example 14. Preparation of tetra-Boc Dab-polymyxin B cycloheptapeptide (INT-14)

Step a. Synthesis of Cbz-L-Dab-I -11

INT-1 1 (2 g, 1 .88 mmol), Z-L-Dab(Boc)-OH-DCHA (1 .20 g, 2.33 mmol), and N-methylmorpholine (0.62 mL, 5.65 mmol) were dissolved in DMF (6 mL) and HATU (0.858 g, 2.25 mmol) was added and the reaction was stirred for 45 minutes. The mixture was applied directly to reversed phase HPLC (25-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were lyophilized to afford 2.1 g of the title compound as a white solid. Yield: 79%. LC/MS (M+2H)/2 = 599.0 (Cbz protected intermediate, loss of 2 Boc groups on LC/MS).

Step b. Preparation of INT-14

The Cbz-L-Dab-INT-12 (2.1 g, 1 .50 mmol) was stirred in methanol (25 mL) in the presence of 5% Pd/C (125 mg) under 1 atm of hydrogen gas for 45 minutes. The mixture was filtered, concentrated and applied to reversed phase HPLC (25-95% acetonitrile in Dl water: 20 minute gradient). The pure fractions were lyophilized to afford 1 .92 g of INT-14 as a white solid. Yield: 96%. LC/MS (M+2H)/2=532.0 (loss of 2 Boc groups on LC/MS). Example 15. Synthesis of INT-15 (tetra-Boc polymyxin E nonapeptide)

Procedu e A

INT-15 was prepared from INT-13 and Z-Thr-OH in a manner similar to that described for INT-16. Yield 73%. LC/MS (M+2H)/2 = 565.8 (loss of 2 Boc groups on LC/MS).

Procedure B

Step a. Enzymatic hydrolysis of colistin to polymyxin E nonapeptide (PMEN)

Colistin sulfate was dissolved in phosphate buffer (1 liter, pH 7, 25 mM), and KCI (1 .5g), EDTA (500 mg), Cysteine-HCI (2g) and Immobilised papain (10 mL, Thermofisher, 16-40 BAEE/mg before immobilisation) was added then the pH was brought back to 7 with potassium phosphate, dibasic. The reaction was shaken at 37 ^SC for 72 hours. The solids were removed by filtration and the aqueous reaction mixture was lyophilized. The white solids were dissolved in 1 N HCI and purified (in 3 aliquots) by reversed phase HPLC (5-50% acetonitrile in Dl water containing 0.1 % TFA: 20 minute gradient). Pure fractions were pooled and lyophilized to afford the product as a white solid. LC/MS 929.5 and 465.0 ((M+2H)/2).

Step b. Selective protection of polymyxin E nonapeptide to tetra-Boc polymyxin E nonapeptide (INT-15)

The title compound may be prepared in a manner analogous to the following literature procedure: O'Dowd, et al. Tetrahedron Letters, Volume 48, Issue 1 1 , 12 March 2007, Pages 2003-2005.

Example 16. Synthesis of INT-16 (tetra-Boc polymyxin B nonapeptide)

Procedu e A

INT-14 (0.22 g, 0.174 mmol), Z-Thr-OH (0.053 g, 0.209 mmol), and N-methylmorpholine (0.057 mL, 5.65 mmol) were dissolved in DMF (6 mL) and HATU (0.070 g, 0.209 mmol) was added and the reaction was stirred for 45 minutes. The mixture was applied directly to reversed phase HPLC (25-95% acetonitrile in Dl water containing 0.1 % formic acid:20 minute gradient). The pure fractions were lyophilized to afford 2.1 g of the title compound as a white solid. LC/MS (M+2H)/2 = 649.4 (Cbz protected intermediate, loss of 2 boc groups on LC/MS). The Cbz protected intermediate was taken up in methanol (10 mL) and stirred in the presence of 5% Pd/C (40 mg) under 1 atmosphere of hydrogen gas for 45 minutes. The mixture was filtered and applied to reversed phase HPLC (20-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were lyophilized to afford 0.187 g of the title compound as a white solid. Yield: 78%, 2 steps. LC/MS (M+2H)/2 = 582.4 (loss of 2 Boc groups on LC/MS).

Procedure B

Tetra-Boc polymyxin B nonapeptide is prepared from polymyxin B via polymyxin B nonapeptide (PMBN) in a manner similar to that described in Example 15, Procedure B.

Example 17. Synthesis of INT-17 (penta-Boc polymyxin E decapeptide)

Penta-Boc polymyxin E decapeptide was prepared analogously to penta-Boc polymyxin B decapeptide (see Example 18), except that polymyxin E was substituted for polymyxin B as the starting material.

Exam -18 (penta-Boc polymyxin B decapeptide)

Step a. Synthesis of Cbz-penta-Boc polymyxin B decapeptide

A solution of INT-1 1 (2.25 g, 2.18 mmol), (2S)-2-[(N-{(2S)-2-{[(benzyloxy)carbonyl]amino}-4-[(tert- butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoic acid (1 .36 g, 2.08 mmol), and DIEA (1 .20 mL, 6.87 mmol), in DMF (5 mL ), were treated with HATU (0.831 g, 2.18 mmol), while stirring at room temperature. After 30 minutes, the product was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 20% to 1 00% methanol and water, using no modifier. Yield 2.50 g, 72% yield, lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 799.5, [(M-2Boc)+2H]/2 = 749.5

Step b. Synthesis of penta-Boc polymyxin B decapeptide

A solution of Cbz-penta-Boc polymyxin B decapeptide (2.5 g, 1 .47 mmol) dissolved in methanol (10 mL), was treated with 5% Pd/C (0.5 g), flushed with hydrogen and stirred for 12 hr. The reaction was filtered through Celite, concentrated, and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 1 .98 g, 86% yield, lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 732.4

Example 19. Synthesis of INT-19 (penta-Boc polymyxin E undecapeptide)

Penta-Boc polymyxin E undecapeptide was prepared analogously to penta-Boc polymyxin B undecapeptide (see Example 20), except that penta-Boc polymyxin E decapeptide was substituted for penta-Boc polymyxin B decapeptide as the starting material. E

Step a. Synthesis of Cbz penta-Boc polymyxin B undecapeptide

A solution of penta-Boc polymyxin B decapeptide (prepared as described in Example 18) (1 .31 g, 0.838 mmol), (2S)-2-{[(benzyloxy)carbonyl]amino}octanoic acid (0.25 g, 0.880 mmol), and DIEA (0.46 mL, 2.64 mmol), in DMF (5 mL ), were treated with HATU (0.334 g, 0.880 mmol), while stirring at room temperature. After 30 minutes, the product was used in the next step without purification, lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 870.0, [(M-2Boc)+2H]/2 = 820.0. Step b. Synthesis of penta-Boc polymyxin B undecapeptide

The solution from Step a was treated with 5% Pd/C (0.5g), flushed with hydrogen and stirred for 2 hr. The reaction was filtered through Celite, concentrated, and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.83 g, 58% yield, lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 803.0, [(M-2Boc)+2H]/2 = 753.0

Example 21. Synthesis of INT-21 (H-Thr-YBoc-Dab-OMe)

Proce

Step a. Preparation of Cbz-Thr-yBoc-Dab-OMe

NH₂-Dab(Boc)-OMe (HCI salt) (5.000g, 1 eq.), Z-NH-L-Thr-OH (5.049g, 1 .05eq.), EDCI (5.350g, 1 .5eq.), HOBt (3.733g, 1 .5eq.) and NaHC0₃ (3.095g, 2eq.) were weighed into a 100- mL round bottom flask. 24 mL of DCM/DMF (4:1 ) was added into the flask. The mixture was stirred at room temperature for about 3 hrs. (TLC or LC/MS monitoring). After completion, EtOAc (200 mL) was added to dilute the reaction mixture and washed with 1 N aq. HCI, saturated NaHC03 and brine. Dried with Na2S04 and condensed. The residue was purified with normal phase silica (2-7% MeOH/DCM) to give 8.24 g pure desired product (>95%).

Step b. Removal of the Cbz Group

Cbz-Thr-yBoc-Dab-OMe (8.24 g) was dissolved in 1 00 mL of MeOH/EtOAc. And 5%Pd/C (3.75 g, 0.1 eq) was added. Under H2 balloon, the reaction mixture was stirred for 2 h. Check TLC and LC/MS to determine completion of the reaction. The reaction was subjected to Celite filtration and with MeOH washing of the solids. Dried to give 5.87 g of free amine (>99%). The material may be used in the next step without purification.

Procedu e B

Z-Thr-OH (2.0 g, 7.9 mmol), methyl (2S)-2-amino-4-[(tert-butoxymethyl)amino]butanoate hydrochloride (2.2 g, 8.7 mmol), HOBt (1 .2 g, 8.7 mmol), EDC (1 .7 g, 8.7 mmol) and DIEA (2.0 g, 1 6 mmol) were stirred in DMF (15 mL) at ambient temperature for 2 hours. The mixture was diluted with ethyl acetate (30 mL) and aqueous 1 N HCI (50 mL). The aqueous phase was extracted with ethyl acetate (3x, 30 mL) and the combined organic extracts were dried over sodium sulfate, filtered and concentrated. The crude dipeptide was stirred in a methanol (50 mL) under 1 atmosphere of hydrogen gas in the presence of 5% Pd/C (200 mg) for 45 minutes. The mixture was filtered, concentrated and purified by reversed phase HPLC (5 to 75% acetonitrile in Dl water containing 0.1 % TFA: 20 minute gradient). The pure fractions were pooled and lyophilized to afford H-Thr-yBoc-Dab-OMe as a white solid. Yield: 82%. LC/MS [M+H⁺]⁺ 334.8.

Example 22. Synthesis of INT-22

Step a. Synthesis of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-

{[(benzyloxy)carbonyl]amino}octanoyl]amino}-4-[(tert-butoxycarbonyl)amino]butanoyl}-L threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate

A solution of methyl (2S)-2-[(N-{(2S)-2-amino-4-[(tert-butoxycarbonyl)amino]butanoyl}-L- threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.600 g, 1 .124 mmol),(2S)-2- {[(benzyloxy)carbonyl]amino}octanoic acid (0.330 g, 1 .124 mmol), and DIEA (0.59 mL, 3.37 mmol) in DMF (4 mL), was treated with HATU (0.470g, 1 .24 mmol), and stirred for 30 min. The reaction was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 10% to 100% acetonitrile and water, using formic acid (0.1 %) as the modifier. Yield 0.726 g, (79%). lon(s) found by LCMS: (M+H)⁺ = 809.5

Step b. Synthesis of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-aminooctanoyl]amino}-4-[(tert- butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate

A mixture of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-{[(benzyloxy)carbonyl]amino}octanoyl]amino}-4- [(tert-butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.300 g, 0.371 mmol), 5% Pd/C (0.150 g), and methanol (4 mL), were charged with hydrogen from a balloon. After 2 hr, the product was filtered through Celite, concentrated, and used in the next step without further purification. Mass recovery 0.250 g, quantitative, lon(s) found by LCMS: (M+H)⁺ = 675.4

Step a. Synthesis of benzyl diethyl 2,2',2"-nitrilotriacetate

Benzyl bromoacetate (4.6 g, 20 mmol) was added into the solution of diethyl 2,2'- azanediyldiacetate (3.8 g, 20 mmol) and DIPEA (3.6 mL, 30 mmol) in 60 mL DMF, the resultant solution was stirred at room temperature for overnight. The reaction solution was concentrated and purified by flash chromatography to provide products. ¹ H NMR (300 MHz, Chloroform-d) δ 7.37 (m, 5H), 5.17 (s, 2H), 4.18 (q, J = 6.9 Hz, 4H), 3.78 (s, 2H), 3.68 (s, 4H), 1 .27 (t, J = 8.2Hz, 6H). LC/MS, 338.2 [M+H]⁺.

Step b. Synthesis of [bis(2-ethoxy-2-oxoethyl)amino]acetic acid

Benzyl diethyl 2,2',2"-nitrilotriacetate (4.6 g, 13.6 mmol) was dissolved into 30 mL MeOH and 30 mL ethyl acetate, then 1 g of 5% palladium on charcoal was added above solution and the mixture was stirred at room temperature under an hydrogen atmosphere overnight. The solids were removed by filtration after completion of the reaction as judged by LCMS. The filtrate was concentrated and used in the next step without any purification. ¹ H NMR (300 MHz, Chloroform-d) δ 4.24 (q, J = 7.1 Hz, 4H), 3.61 (s, 4H), 3.53 (d, J = 10.1 Hz, 2H), 1 .31 (t, J = 7.2 Hz, 6H). LC/MS, 247.2 [M+H]⁺ .

Step c. Synthesis of diethyl 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)diacetate

To the solution of [bis(2-ethoxy-2-oxoethyl)amino]acetic acid (4.4 g, 18 mmol), 2-(2- aminoethoxy)ethyl 6-deoxy-alpha-L-mannopyranoside (4.51 g, 18 mmol) in DMF (50 mL) was added EDC (4 g, 20 mmol), HOBT (2.7g, 20 mmol), Hunig's base (4.2 mL, 30 mmol) at room temperature. The solution was stirred for overnight. The resultant solution was removed DMF and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, 7.0 g, 81 % yield. ¹ H NMR (300 MHz, methanol-ck) δ 4.19 (qd, J = 7.2, 3.0 Hz, 2H), 3.88 - 3.55 (m, 4H), 3.54 - 3.33 (m, 3H), 2.94 (s, 1 H), 2.83 (s, 1 H), 2.36 (t, J = 8.1 Hz, OH), 2.03 (p, J = 7.7 Hz, OH), 1 .43 - 1 .21 (m, 4H). LC/MS 481 .2 [M+H]⁺

Step d. Synthesis of 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}azanediyl)diacetic acid

Lithium hydroxide (20 mmol) in 10 mL H2O was added to the solution of diethyl 2,2'-({2-

[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)diacetate (4.80g, 10 mmol) in mix solvent of 10 mL H2O, 20 mL MeOH and 20 mL THF. The resultant solution was stirred for 1 hours at room temperature. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. 5.7g, yield of 92%. ¹ H NMR (300 MHz, methanol-ck) δ 4.73 (s, 1 H), 3.80 (dd, J = 15.5, 5.5 Hz, 2H), 3.73 - 3.61 (m, 7H), 3.60 (d, J = 10.6 Hz, 1 H), 3.59 (s, 6H), 3.40 (d, J = 23.5 Hz, 7H), 1 .27 (d, J = 6.3 Hz, 3H). LC/MS 425.2 [M+H]⁺ Step e. Synthesis of dibenzyl 14-{2-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}-4,12,16,24-tetraoxo-8,20-dioxa- 5,11 ,14,17,23-pentaazaheptacosane-1 ,27-dioate

To the solution 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}azanediyl)diacetic acid (Example 23, Step d) (1 .3 g, 3 mmol) and 4-(benzyloxy)-4-oxobutanoic acid (1 .8 g, 6.1 mmol) in DMF (30 ml_) was added EDC (1 .4g, 7 mmol), HOBT (1 .0g, 7 mmol), Hunig's base (1 .4 ml_, 10 mmol) at room temperature. The solution was stirred for overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 1 00% acetonitrile and water, using 0.1 %

trifluoroacetic acid as the modifier. The product was obtained as an oil, 2.2 g, 75% yield. ¹ H NMR (300 MHz, methanol-dt) δ 7.36 (s, 10H), 5.13 (s, 4H), 4.72 (s, 1 H), 3.81 (s, 2H), 3.70 - 3.46 (m, 16H), 3.46 - 3.24 (m, 85H), 2.70 (q, J = 6.9, 5.4 Hz, 5H), 2.55 (d, J = 7.0 Hz, 3H), 1 .27 (d, J = 6.2 Hz, 3H). LC/MS 977.5 [M+H]⁺ Step f. Synthesis of 14-{2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}-4,12,16,24-tetraoxo-8,20-dioxa-5,11 ,14,17,23-pentaazaheptacosane-1 ,27-dioic acid

Dibenzyl 14-{2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}- 4,12,16,24-tetraoxo-8,20-dioxa-5,1 1 ,14,17,23-pentaazaheptacosane-1 ,27-dioate (816mg, 0.836 mmol) was dissolved into 5 mL MeOH and 5 mL ethyl acetate, then 0.5 g of 5% Palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for overnight. The palladium charcoal was removed by filtration after completed the reaction by LCMS. The filtrate was concentrated and used next step without any purification. ¹ H NMR (300 MHz, methanol- dt) 5 4.73(s, 1 H) 3.74 (s, 7H), 3.70 - 3.49 (m, 15H), 3.43 (dd, J = 14.3, 7.8 Hz, 8H), 3.36 (s, 48H), 2.69 - 2.43 (m, 8H), 1 .27 (d, J = 6.2 Hz, 3H). LC/MS, 797.4 [M+H]⁺ .

Step g. Synthesis of per-Boc-(Compound 1)

To the solution of 14-{2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}-4,12,16,24-tetraoxo-8,20-dioxa-5,1 1 ,14,17,23-pentaazaheptacosane-1 ,27-dioic acid (80 mg, 0.10 mmol), triethylamine (0.07 mL, 0.5 mmol) and INT-16 (270 mg, 0.2 mmol) in 5 mL DMF was added HATU (78 mg, 0.2 mmol). The reaction was stirred for 1 hr and the resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of product, tri-tert-butyl {[(2S,5R,8S,1 1 S,14S,17S,22S)-5-benzyl-22-{[(8S,1 1 S,41 S,44S)-8- {[(3S,6S,9S,12S,15R,18S,21 S)-15-benzyl-6,9,18-tris{2-[(tert-butoxycarbonyl)amino]ethyl}-3-[(1 R)-1 - hydroxyethyl]-12-(2-methylpropyl)-2,5,8,1 1 ,14,17,20-heptaoxo-1 ,4,7,10,13,16,19-heptaazacyclotricosan- 21 -yl]carbamoyl}-44-{2-[(tert-butoxycarbonyl)amino]ethyl}-1 1 ,41 -bis[(1 R)-1 -hydroxyethyl]-2,2-dimethyl- 4,10,13,16,24,28,36,39,42,45-decaoxo-26-(2-oxo-2-{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyloxan-2-yl]oxy}ethoxy)ethyl]amino}ethyl)-3,20,32-trioxa-5,9,12,17,23,26,29,35,40,43- decaazapentatetracontan-45-yl]amino}-17-[(1 R)-1 -hydroxyethyl]-8-(2-methylpropyl)-3,6,9,12,1 5,18,23- heptaoxo-1 ,4,7,10,13,16,19-heptaazacyclotricosane-2,1 1 ,14-triyl]tri(ethane-2,1 -diyl)}triscarbamate. 220 mg, 65% yield.

Step h. Synthesis of Compound 1

The per-Boc-(Compound 1 ) from Step g was treated in 2 ml_ DCM and 2 ml_ TFA at room temperature, the solution was stirred 10 min, then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield of (2S,5S,35S,38S)-2,38-bis(2-aminoethyl)- 5,35-bis[(1 R)-1 -hydroxyethyl]-4,7,10,18,22,30,33,36-octaoxo-20-(2-oxo-2-{[2-(2-{[(2R,3R,4R,5R,6S)- 3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}ethoxy)ethyl]amino}ethyl)-N~1 ~,N~39~- bis(3S,6S,9S,12S,15R,18S,21 S)[6,9,18-tris(2-aminoethyl)-1 5-benzyl-3-[(1 R)-1 -hydroxyethyl]-12-(2- methylpropyl)-2,5,8,1 1 ,14,17,20-heptaoxo-1 ,4,7,1 0,13,16,1 9-heptaazacyclotricosan-21 -yl]-14,26-dioxa- 3,6,1 1 ,17,20,23,29,34,37-nonaazanonatriacontane-1 ,39-diamide was 0.120 g, 67% yield, lon(s) found by LCMS: (M+2H)/2 = 1343.7, (M+3H)/3 = 896.2, (M+4H)/4 =672.4, (M+5H)/5=538.1 .

Example 2

The title compound was prepared analogously to Compound 1 except INT-15 was substituted for INT-16 in the sequence, lon(s) found by LCMS (M+2H)/2 = 1309.8, (M+3H)/3 =873.5, (M+4H)/4 =655.4. (M+5H)/5 =524.5.

Exampl

The title compound was prepared analogously to Compound 10. lon(s) found by LCMS: (M+2H)/2 = 1480.8. (M+3H)/3 = 987.5, (M+4H)/4 =740.9, (M+5H)/5 =592.9. (M+6H)/6 =494.3. Exam

The title compound was prepared analogously to Compound 2. lon(s) found by LCMS: (M+2H)/2 = 1445.8, (M+3H)/3 = 964.9. (M+4H)/4 =724.0, (M+5H)/5 =579.3.

Example 27. Preparation of Compound 5

Step a. Preparation of benzyl {21-[(6-deoxy-alpha-L-mannopyranosyl)oxy]-15-oxo-3,6,9,12,19- pentaoxa-16-azahenicosan-1-yl}carbamate

2-(2-aminoethoxy)ethyl 6-deoxy-alpha-L-mannopyranoside (500 mg, 1 .25 mmol), 3-oxo-1 -phenyl- 2,7,10,13,16-pentaoxa-4-azanonadecan-19-oic acid (325 mg, 1 .29 mmol), EDC (287 mg, 1 .50 mmol) and DIEA (193 mg, 1 .50 mmol) were stirred together in DMF (1 .5 mL) for 3 hours. The mixture was applied directly to reversed phase HPLC (20-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and lyophilized to afford benzyl {21 -[(6-deoxy-alpha-L- mannopyranosyl)oxy]-1 5-oxo-3,6,9,12,19-pentaoxa-16-azahenicosan-1 -yl}carbamate as a clear oil. Yield: 91 %. LC/MS [m+H]⁺: 633.6.

Step b. Synthesis of 1-[(6-deoxy-alpha-L-mannopyranosyl)oxy]-7,23-dioxo-3,10,13,16,19- pentaoxa-6,22-diazahexacosan-26-oic acid

Benzyl {21 -[(6-deoxy-alpha-L-mannopyranosyl)oxy]-15-oxo-3,6,9,12,19-pentaoxa-16- azahenicosan-1 -yl}carbamate (250 mg, 0.395 mmol) was stirred under 1 atmosphere of hydrogen gas i the presence of 5% Pd/C (70 mg) at room temperature for 2 hours. The mixture was filtered,

concentrated, and taken up in methanol (4 mL) and succinic anhydride (40 mg, 0.395 mmol) and trimethylamine (40 mg, 0.395 mmol) were added. The reaction was stirred at room temperature for 2 hours and then concentrated, taken up in Dl water (2 mL) and applied to reversed phase HPLC (5-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and lyophilized to afford 1 -[(6-deoxy-alpha-L-mannopyranosyl)oxy]-7,23-dioxo-3,1 0,13,16,19-pentaoxa- 6,22-diazahexacosan-26-oic acid as a clear, viscous oil. Yield: 85%. LC/MS [M-H-] = 597.4.

Step c. Sy

HATU (1 12 mg, 0.250 mmol) was added to stirring mixture of tri-tert-butyl

{[(2S,5R,8S,1 1 S,14S,17S,22S)-22-({(2S,5S,24S,27S)-29-amino-2-(2-aminoethyl)-27-

{[(3S,6S,9S,12S,15R,18S,21 S)-15-benzyl-6,9,18-tris{2-[(tert-butoxycarbonyl)amino]ethyl}-3-[(1 R)-1 - hydroxyethyl]-12-(2-methylpropyl)-2,5,8,1 1 ,14,17,20-heptaoxo-1 ,4,7,10,13,16,19-heptaazacyclotricosan- 21 -yl]carbamoyl}-5,24-bis[(1 R)-1 -hydroxyethyl]-4,7,22,25-tetraoxo-10,13,1 6,1 9-tetraoxa-3, 6,23,26- tetraazanonacosanan-1 -oyl}amino)-5-benzyl-17-[(1 R)-1 -hydroxyethyl]-8-(2-methylpropyl)- 3,6,9,12,15,18,23-heptaoxo-1 ,4,7,10,13,16,19-heptaazacyclotricosane-2,1 1 ,14-triyl]tri(ethane-2,1 - diyl)}triscarbamate (279 mg, 0.1 00 mmol), 1 -[(6-deoxy-alpha-L-mannopyranosyl)oxy]-7,23-dioxo- 3,10,13,16,19-pentaoxa-6,22-diazahexacosan-26-oic acid (150 mg, 0.250 mmol), and DIEA (77 mg, 0.601 mmol) in DMF (4 mL). The reaction was stirred at room temperature for 4 hours then applied directly to reversed phase HPLC (25-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and concentrated. The Boc-protected intermediate was stirred in a 1 /1 mixture of TFA/DCM containing 0.1 % thioanisole (5 mL) at room temperature for 30 minutes. The solvent was removed by rotary evaporation and the crude product was purified by to reversed phase HPLC (0-35% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and lyophilized to afford (23S,26S,45S,48S)-26,45-bis[(1 R)-1 -hydroxyethyl]- 16,1 9,25,28,43,46, 52, 55-octaoxo-N~1 ~,N~70~-bis[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyloxan-2-yl]oxy}ethoxy)ethyl]-N~23~,N~48~-bis(3S,6S,9S,12S,15R,18S,21 S)[6,9,18-tris(2- aminoethyl)-15-benzyl-3-[(1 R)-1 -hydroxyethyl]-12-(2-methylpropyl)-2,5,8,1 1 ,14,17,20-heptaoxo- 1 , 4,7,10,13, 16,1 9-heptaazacyclotricosan-21 -yl]-3, 6, 9, 12,31 , 34,37,40,59,62,65, 68-dodecaoxa- 15,20,24,27,44,47,51 ,56-octaazaheptacontane-1 ,23,48,70-tetracarboxamide (Compound 5) as a white solid. Yield: 10%, 2 steps. LC/MS (M+4H)/4 = 837.0.

Example 28. Synthesis of Compound 6

Step a. Synthesis of benzyl bis(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethyl)carbamate

To the solution rhamnose(2R,3R,4R,5R,6S)-2-(2-(2-aminoethoxy)ethoxy)-6-methyltetrahydro-2H- pyran-3,4,5-triol (INT-1 ) (1 .0 mmol) and 2,2'-(((benzyloxy)carbonyl)azanediyl)diacetic acid (0.50 g, 2 mmol) in DMF (1 0 mL) was added EDC (1 .0 g, 5 mmol), HOBT (750 mg, 5 mmol) and Hunig's base (1 .4 mL, 10 mmol) at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil 620 mg, 48% yield. LC/MS 734.3 [M+H]⁺

Step b. Synthesis of ethyl 6-(2-ethoxy-2-oxoethyl)-4,8,11-trioxo-9-(2-oxo-2-((2-(2- (((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)ethoxy)ethyl)amino)ethyl)-17-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)-15-oxa-3,6,9,12-tetraazaheptadecan-1 -oate

Benzyl bis(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)ethoxy)ethyl)amino)ethyl)carbamate (400 mg, 0.05 mmol) was dissolved into 5 mL MeOH and 5 ml_ ethyl acetate, then 100 mg of 5% palladium on charcoal was added to the above solution and the mixture was stirred at room temperature under a hydrogen atmosphere for overnight. The palladium on charcoal was removed by filtration after completion of the reaction by LCMS. The filtrate was

concentrated and used for next step without any purification. LC/MS [M]+1 600.3.

Step c. Synthesis of ethyl 12-(2-ethoxy-2-oxoethyl)-7,10-dioxo-9-(2-oxo-2-{[2-(2-

{[(2/?,3/?,4/?,5/?,6¾-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}ethoxy)ethyl]amino}ethyl)-1-

{[(2/?,3/?,4/?,5/?,6¾-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-3-oxa-6,9,12-triazatetradecan-14-oate

The amine product from the previous step was dissolved into 5 mL DMF, then 2-(bis(2-ethoxy-2- oxoethyl)amino)acetic acid (240 mg, 1 mmol), EDC (1200 mg, 10 mmol), HOBT (150 mg, 5 mmol),

Hunig's base (0.28 mL, 2 mmol) were added to the solution at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil 320 mg, 75% yield. LC/MS [M+H]⁺ 829.4

Step d. Synthesis of 12-(carboxymethyl)-7,10-dioxo-9-(2-oxo-2-{[2-(2-{[(2/?,3/?,4/?,5/?,6S)-3,4,5- trihydroxy-6-methyloxan-2-yl]oxy}ethoxy)ethyl]amino}ethyl)-1-{[(2/?,3/?,4/?,5/?,6¾-3,4,5-trihydroxy- 6-methyloxan-2-yl]oxy}-3-oxa-6,9,12-triazatetradecan-14-oic acid

Lithium hydroxide (24 mg, 1 mmol) in 3 mL H2O was added to the solution of ethyl 6-(2-ethoxy-2- oxoethyl)-4,8,1 1 -trioxo-9-(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)ethoxy)ethyl)amino)ethyl)-17-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)-15-oxa-3,6,9,12-tetraazaheptadecan-1 -oate (200 mg, 0.25 mmol) in a mixed solvent of 3 mL MeOH and 3 mL THF. The resultant solution was stirred for 1 hour at room temperature then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, 180 mg 90% yield. LC/MS [M+H]⁺ 773.2.

Step e. Preparation of dibenzyl 14-(2-(bis(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethyl)amino)-2-oxoethyl)-4,12,16,24- tetraoxo-8,20-dioxa-5,11 ,14,17,23-pentaazaheptacosane-1 ,27-dioate

12-(carboxymethyl)-7,10-dioxo-9-(2-oxo-2-{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-ethyloxan -2-yl]oxy}ethoxy)ethyl]amino}ethyl)-1 -{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-3-oxa- 6,9,12-triazatetradecan-14-oic acid (150 mg, 0.2 mmol), benzyl 4-{[2-(2-aminoethoxy)ethyl]amino}-4- oxobutanoate (100 mg, 0.34 mmol), EDC (160 mg, 0.8 mmol), HOBt (120 mg, 0.8 mmol) and TEA(0.14 mL, 1 mmol) were dissolved into 5 mL DMF and the solution was stirred for overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil 120 mg, 62% yield. LC/MS [M+H]⁺ 1325.6. Step f. Synthesis of 14-(2-(bis(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethyl)amino)-2-oxoethyl)-4,12,16,24- tetraoxo-8,20-dioxa-5, 11 ,14,17,23-pentaazaheptacosane-1 ,27-dioic acid

Dibenzyl 14-(2-(bis(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)ethoxy)ethyl)amino)ethyl)amino)-2-oxoethyl)-4,12,16,24-tetraoxo-8,20-dioxa-

5,1 1 ,14,17, 23-pentaazaheptacosane-1 ,27-dioate (100 mg, 0.08 mmol) was dissolved into 3 mL MeOH and 3 mL ethyl acetate, then 30 mg of 5% palladium on charcoal was added above solution and the mixture was stirred at room temperature under a hydrogen atmosphere overnight. The solids were removed by filtration after completion of the reaction as evidenced by LCMS. The filtrate was

concentrated and used for next step without any purification. LC/MS [M+H]⁺ 1 145.5.

Step

The title compound was prepared analogously to the procedure described for Compound 1 (Steps g and h) except 14-(2-(bis(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro- 2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethyl)amino)-2-oxoethyl)-4,12,1 6,24-tetraoxo-8,20-dioxa- 5,1 1 ,14,17,23-pentaazaheptacosane-1 ,27-dioic acid (Example 28, Step f) was used as the linker, lon(s) found by LCMS (M+3H)/3 =1012.2, (M+4H)/4 =759.4. (M+5H)/5 =607.7. (M+6H)/6 =506.6.

Example 2

The title compound was prepared analogously to Compound 10. lon(s) found by LCMS (M+3H)/3 = 1 037.2, (M+4H)/4 =778.2, (M+5H)/5 =622.7. (M+6H)/6 =518.1 . Example 30.

Step a. Synthesis of di-(4-formylphenyl)methyl amino-PEG10-acid

Terephthalaldehyde (536.4 mg, 4.0 mmol) and amino-PEG1 0-acid (529.6 mg, 1 mmol) were dissolved in anhydrous EtOH (8 mL) by gently heating with a heat gun. After cooling to room temperature, sodium triacetoxyborohydride (635.8 mg, 3 mmol) was added in two portions over 30 minutes to the solution. The mixture was stirred for 2 hours and directly purified by RPLC (0 to 100% acetonitrile in water). Yield 289 mg, 37.7 %. lon(s) found by LCMS: [M + H]⁺ = 766.

Step b. Synthesis of N-(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl) di-(4- formylphenyl) Compound 8 precursor

To a solution of a mixture of di-(4-formylphenyl)methyl amino-PEG10-acid (120 mg, 0.157 mmol) and HATU (71 .5 mg, 0.188 mmol) in anhydrous DMF (0.5 mL) was added DIPEA (40 mg, 0.316 mmol). After the solution was stirred for 10 minutes, it was added with 2-(2-aminoethoxy)ethyl 6-deoxy-alpha-L- altropyranoside TFA salt (59.2 mg, 0.236 mmol). The resulting mixture was stirred for 30 minutes, then directly purified through RPLC (0 to 40% acetonitrile and water). Yield 1 10 mg, 70.1 %. lon(s) found by LCMS calculated: (M+2H)/2 = 500. Step c. Synthesis of 4-Z-1-Boc-piperazine-INT-14

A mixture of (R)-4-(benzyloxy)carbonyl)-1 -(tert-butoxycarbonyl)piperazine-2-carboxylic acid (1 75 mg, 0.48 mmol) and HATU (190 mg, 0.5 mmol) was dissolved in anhydrous DMF (1 mL), and the solution was treated with DIPEA (130 mg, 1 mmol). After the mixture was stirred for 10 minutes, it was added with INT-14 (505 mg, 0.4 mmol). The resulting mixture was stirred for 1 hour, then purified through RPLC (25 to 100 % acetonitrile and water, using 0.1 % formic acid as modifier). Yield 572.4 mg, 94.9 %. lon(s) found by LCMS: [(M - 2Boc)/2]+1 = 705. Step d. Synthesis of 1-Boc-piperazine-INT-14

4-Z-1 -Boc-piperazine-INT-14 (522.9 mg, 0.325 mmol) was dissolved in MeOH (20 mL) and treated with Pd/C. The suspension was stirred under hydrogen for 1 .5 hours. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation to afford the title product as a tan solid. Yield 465 mg, 97%. lon(s) found by LCMS calculated: [(M-Boc)/2]+1 = 688.

Step e. Synthesis of Compound 8

A mixture of 1 -Boc-piperazine-INT-14 (98 mg, 0.0644 mmol) and N-(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl) di-(4-formylphenyl) Compound 8 precursor (32 mg, 0.032 mmol) from Step b was dissolved in anhydrous EtOH (-2 mL). NaBH(OAc)3 (41 mg) was added, and the resulting mixture was stirred overnight. Four additional portions of NaBH(OAc)3 (41 mg x 4) were added every 1 hour. Stirring of the reaction was continued for another 2 hours, then purified through HPLC (25 to 50 % acetonitrile and water, using 0.1 % formic acid as modifier). The collected fractions were lyophilized to a white solid (1 9.7 mg). The materials were treated with TFA (-0.5 mL), and the solution was stirred for 20 minutes. The mixture was purified by HPLC (0 to 8.5 % acetonitrile and water, using 0.1 % formic acid). lon(s) found by LCMS: (M+5H)/5 = 583.

Step a. Synthesis of (S)-benzyl 3-((tert-butoxycarbonyl)amino)-4-(octylamino)-4-oxobutanoate

To a solution of n-octylamine (650 mg, 5 mmol) and (S)-4-(benzyloxy)-2-((tert- butoxycarbonyl)amino)-4-oxobutanoic acid (1 .65 g, 5 mmol) in DMF (30 mL) was added EDC (1 .2 g, 6 mmol), HOBT (900 mg, 6 mmol), Hunig's base (1 .4 mL, 10 mmol) at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil 2.1 g, 85% yield. LC/MS 435.3 [M+H]⁺ Step b. Synthesis of (S)-diethyl 2,2'-((2-((4-(benzyloxy)-1-(octylamino)-1 ,4-dioxobutan-2-yl)amino)- 2-oxoethyl)azanediyl)diacetate

(S)-benzyl 3-((tert-butoxycarbonyl)amino)-4-(octylamino)-4-oxobutanoate (440 mg, 1 mmol) was treated with TFA and stirred for half hour and then concentrated and dried under high vacuum used for next step without purification.

The deprotected product from the previous step was mixed with [bis(2-ethoxy-2- oxoethyl)amino]acetic acid (250 mg, 1 mmol) in DMF (10 mL) was added EDC (250 mg, 1 .25 mmol), HOBT (200 mg, 1 .25 mmol), Hunig's base (0.3 mL, 2 mmol) at room temperature. The solution was stirred overnight. The solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil 450 mg, 85% yield. LC/MS 564.3 [M+H]⁺

Step c. Synthesis of (S)-ethyl 3-(2-ethoxy-2-oxoethyl)-7-(octylcarbamoyl)-5,9-dioxo-15- (((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-13-oxa-3,6,10- triazapentadecan-1-oate

(S)-diethyl 2,2'-((2-((4-(benzyloxy)-1 -(octylamino)-l ,4-dioxobutan-2-yl)amino)-2- oxoethyl)azanediyl)diacetate (300 mg, 0.5 mmol) was dissolved into 3 mL MeOH and 3 mL ethyl acetate, then 0.1 g of 5% palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere overnight. The palladium on charcoal was removed by filtration after completion of the reaction by LCMS. The filtrate was concentrated and used in the next step without any purification. LCMS [M+H]+ calculated 473.3.

To the above debenzylated products and 2-(2-aminoethoxy) ethyl 6-deoxy-alpha-L-mannopyranoside (INT-1 ) (150 mg, 0.6 mmol) in DMF (5 mL) was added EDC (120 mg, 0.6 mmol), HOBT (100 mg, 0.6 mmol) and Hunig's base (0.14 mL, 1 mmol) at room temperature. The solution was stirred overnight. The mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 1 00% acetonitrile and water, using 0.1 %

trifluoroacetic acid as the modifier. The product was obtained as an oil, LC/MS 707.4 [M+H]⁺ Step d. Synthesis of (S)-3-(carboxymethyl)-7-(octylcarbamoyl)-5,9-dioxo-15-(((2R,3R,4R,5R,6S)- 3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-13-oxa-3,6,10-triazapentadecan-1-oic acid

Lithium hydroxide (24 mg, 1 mmol) in 3 mL H2O was added to a solution of (S)-ethyl 3-(2-ethoxy- 2-oxoethyl)-7-(octylcarbamoyl)-5,9-dioxo-15-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)-13-oxa-3,6,10-triazapentadecan-1 -oate (210 mg, 0.3 mmol) in a mixed solvent of 3 mL MeOH and 3 mL THF. The resultant solution was stirred for 1 hour at room temperature. The reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 1 00% acetonitrile and water, using 0.1 %

trifluoroacetic acid as the modifier. The product was obtained as an oil, 180 mg 85% yield. LC/MS [M+H] 651 .3.

Step e. Synthesis of dibenzyl 14-(2-(((S)-1-(octylamino)-1 ,4-dioxo-4-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5- trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)butan-2-yl)amino)-2- oxoethyl)-4,12,16,24-tetraoxo-8,20-dioxa-5,11 ,14,17,23-pentaazaheptacosane-1 ,27-dioate

The above (S)-3-(carboxymethyl)-7-(octylcarbamoyl)-5,9-dioxo-1 5-(((2R,3R,4R,5R,6S)-3,4,5- trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-13-oxa-3,6,10-triazapentadecan-1 -oic acid (130 mg, 0.2 mmol) and benzyl 4-((2-(2-aminoethoxy)ethyl)amino)-4-oxobutanoate (150 mg, 0.6 mmol) in DMF (5 mL) was added EDC (120 mg, 0.6 mmol), HOBT (100 mg, 0.6 mmol), Hunig's base (0.14 mL, 1 mmol) at room temperature. The solution was stirred overnight. The mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluting with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, LC/MS 602.3 [M+2H]/2

Step f. Synthesis of 14-(2-(((S)-1-(octylamino)-1 ,4-dioxo-4-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5- trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)butan-2-yl)amino)-2- oxoethyl)-4,12,16,24-tetraoxo-8,20-dioxa-5,11 ,14,17,23-pentaazaheptacosane-1 ,27-dioic acid

Dibenzyl 14-(2-(((S)-1 -(octylamino)-l ,4-dioxo-4-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)butan-2-yl)amino)-2-oxoethyl)-4,12,16,24- tetraoxo-8,20-dioxa-5,1 1 ,14,17,23-pentaazaheptacosane-1 ,27-dioate..(120 mg, 0.1 mmol) was dissolved into 3 mL MeOH and 3 mL ethyl acetate. Then 0.1 g of 5% palladium on charcoal was added to the solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for overnight. The palladium on charcoal was removed by filtration after completion of the reaction as determined by LCMS. The filtrate was concentrated and used in the next step without any purification. LCMS: [M+2H]/2 512.3.

Step g. Preparation of Compound 9

The title compound was prepared analogously to Compound 1 utilizing 14-(2-(((S)-1 -(octylamino)- 1 ,4-dioxo-4-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)ethoxy)ethyl)amino)butan-2-yl)amino)-2-oxoethyl)-4,12,16,24-tetraoxo-8,20-dioxa-5,1 1 ,14,17,23- pentaazaheptacosane-1 ,27-dioic acid (Example 31 , Step f) as the linker, lon(s) found by LCMS: (M+2H)/2 = 1456.8, (M+3H)/3 = 970.6, (M+4H)/4 =728.90, (M+5H)/5 =583.3.

Example 32. Synthesis of Compound 10

Step a. Synthesis of 6-(2-ethoxy-2-oxoethyl)-4,8-dioxo-

3,12,15,18,21 ,24,27,30,33,36,39,42,45,48,51 ,54,57,60,63,66,69,72,75,78,81 -pentacosaoxa-6,9- diazatetraoctacontan-84-oic acid

To a solution of [bis(2-ethoxy-2-oxoethyl)amino]acetic acid (62 mg, 25 mmol and amino-PEG24-t- butyl ester (600 mg, 0.5 mmol ) in DMF (5 mL) was added EDC (120 mg, 0.6 mmol), HOBT (100 mg, 6 mmol) and Hunig's base (0.3 mL, 2 mmol) at room temperature. The solution was stirred overnight. The DMF was removed under reduced pressure and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier.

The above product was treated with TFA and stirred for 30 min, concentrated and dried under high vacuum and used directly for the next step without purification. LC/MS [M/2-t-Butyl]+1 688.4.

Step b. Synthesis of ethyl 3-(2-ethoxy-2-oxoethyl)-5,81-dioxo-87-(((2R,3R,4R,5R,6S)-3,4,5- trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)- 9,12,15,18,21 ,24,27,30,33,36,39,42,45,48,51 ,54,57,60,63,66,69,72,75,78,85-pentacosaoxa-3,6,82- triazaheptaoctacontan-1 -oate

To a solution of 6-(2-ethoxy-2-oxoethyl)-4,8-dioxo- 3,12,15,18,21 ,24,27,30,33,36,39,42,45,48,51 ,54,57,60,63,66,69,72,75,78,81 -pentacosaoxa-6,9- diazatetraoctacontan-84-oic acid (270 mg, 0.2 mmol) and 2-(2-aminoethoxy)ethyl 6-deoxy-alpha-L- mannopyranoside (75 mg, 0.3 mmol) in DMF (5 mL) was added EDC (70 mg, 0.35 mmol), HOBT (60 mg, 0.4 mmol) and Hunig's base (0.3 mL, 2 mmol) at room temperature. The solution was stirred overnight. The reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, 150 mg 75% yield. LC/MS (M+2H)/2 805.

Step c. Synthesis of 3-(carboxymethyl)-5,81-dioxo-87-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-

9,12,15,18,21 ,24,27,30,33,36,39,42,45,48,51 ,54,57,60,63,66,69,72,75,78,85-pentacosaoxa-3,6,82- triazaheptaoctacontan-1-oic acid

Lithium hydroxide (12 mg, 0.5 mmol) in 3 mL H2O was added to the solution of 3-(2-ethoxy-2- oxoethyl)-5,81 -dioxo-87-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)- 9, 12, 15, 18,21 ,24,27,30,33,36,39,42,45,48, 51 ,54,57,60,63, 66, 69, 72, 75,78, 85-pentacosaoxa-3, 6,82- triazaheptaoctacontan-1 -oate (150 mg, 0.1 mmol) in mix solvent of 3 mL MeOH and 3 mL THF. The resultant solution was stirred for 1 hour at room temperature. The reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, 120 mg 75% yield. LC/MS (M+2H)/2 776.9. Step d. Synthesis of dibenzyl 14-(2,78-dioxo-84-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-

6,9,12,15,18,21 , 24,27,30,33,36,39,42,45,48,51 , 54,57,60,63,66,69,72,75,82-pentacosaoxa-3,79- diazatetraoctacontyl)-4,12,16,24-tetraoxo-8,20-dioxa-5,11 ,14,17,23-pentaazaheptacosane-1 ,27- dioate

To a solution 3-(carboxymethyl)-5,81 -dioxo-87-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)-

9, 12, 15, 18,21 ,24,27,30,33,36,39,42,45,48, 51 ,54,57,60,63, 66, 69, 72, 75,78, 85-pentacosaoxa-3, 6,82- triazaheptaoctacontan-1 -oic acid (150 mg, 0.1 mmol) and benzyl 4-((2-(2-aminoethoxy)ethyl)amino)-4- oxobutanoate (1 00 mg, 0.3 mmol) in DMF (33 mL) was added EDC (60 mg, 0.3 mmol), HOBT (50 mg, 0.3 mmol) and Hunig's base (0.14 mL, 1 mmol) at room temperature. The solution was stirred overnight. The reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, LC/MS 1053.1 [M/2]+H⁺

Step e. Synthesis of 14-(2,78-dioxo-84-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)-6,9,12,15,18,21 , 24,27,30,33,36,39,42,45,48,51 , 54,57,60,63,66,69,72,75,82- pentacosaoxa-3,79-diazatetraoctacontyl)-4,12,16,24-tetraoxo-8,20-dioxa-5,11 ,14,17,23- pentaazaheptacosane-1 ,27-dioic acid

Dibenzyl 14-(2,78-dioxo-84-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)-6,9,12,1 5,1 8,21 ,24,27,30,33,36,39,42,45,48,51 ,54,57,60,63,66, 69, 72,75, 82-pentacosaoxa-3, 79- diazatetraoctacontyl)-4,12,16,24-tetraoxo-8,20-dioxa-5,1 1 ,14,17,23-pentaazaheptacosane-1 ,27-dioate (1 10 mg, 0.05 mmol) was dissolved into 3 mL MeOH and 3 mL ethyl acetate, then 0.1 g of 5% palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for overnight. The palladium on charcoal was removed by filtration after the reaction was determined to reach completion by LCMS. The filtrate was concentrated and used next step without any purification. LCMS: [M+2H]/2 963.0

Step f. Preparation of Compound 10

The title compound was prepared analogously using the procedure described in Example 23

(Steps g and h) but utilized 14-(2,78-dioxo-84-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro- 2H-pyran-2-yl)oxy)-6, 9, 12, 15, 18,21 ,24,27,30,33,36,39,42,45,48, 51 ,54,57,60,63,66, 69,72,75,82- pentacosaoxa-3,79-diazatetraoctacontyl)-4,12,16,24-tetraoxo-8,20-dioxa-5,1 1 ,14,17,23- pentaazaheptacosane-1 ,27-dioic acid (Example 32, Step e) as the linker, lon(s) found by LCMS:

(M+3H)/3 = 1272.1 , (M+4H)/4 =954.3, (M+5H)/5 =763.6. (M+6H)/6 =636.5. Example 33. Synthesis of Compound 11

Step a. Synthesis of dimethyl (2S,5S,8S,16S,19S,22S)-12-[(benzyloxy)carbonyl]-2,8,16,22- tetrakis{2-[(tert-butoxycarbonyl)amino]ethyl}-5,19-bis[(1 R)-1-hydroxyethyl]-4,7,10,14,17,20- hexaoxo-3,6,9,12,15,18,21 -heptaazatricosane-1 ,23-dioate

A stirring solution of methyl (2S)-2-[(N-{(2S)-2-amino-4-[(tert-butoxycarbonyl)amino]butanoyl}-L- threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.399 g, 0.748 mmol), 2,2'- {[(benzyloxy)carbonyl]azanediyl}diacetic acid (0.100 g, 0.374 mmol), and DIEA(0.261 mL, 1 .50 mmol), in DMF (1 mL), were treated with a solution of HATU (0.285 g, 0.748 mmol) added dropwise over 30 minutes, at room temperature. The desired product was isolated by reversed phase liquid

chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.200 g, 41 %. lon(s) found by LCMS: (M+H)+ = 1298.7 Step b. Synthesis of (2S,5S,8S,16S,19S,22S)-12-[(benzyloxy)carbonyl]-2,8,16,22-tetrakis{2-[(tert- butoxycarbonyl)amino]ethyl}-5,19-bis[(1 R)-1-hydroxyethyl]-4,7,10,14,17,20-hexaoxo- 3,6,9,12,15,18,21 -heptaazatricosane-1 ,23-dioic acid

A solution of dimethyl (2S,5S,8S,1 6S,19S,22S)-12-[(benzyloxy)carbonyl]-2,8,16,22-tetrakis{2- [(tert-butoxycarbonyl)amino]ethyl}-5,19-bis[(1 R)-1 -hydroxyethyl]-4,7,10,14,17,20-hexaoxo- 3,6, 9, 12, 15, 18,21 -heptaazatricosane-1 ,23-dioate (0.380 g, 0.242 mmol), in methanol (1 mL) was treated with a solution of lithium hydroxide (0.028 g, 1 .1 71 mmol), in water (1 mL), then stirred at room temperature for 30 minutes. The reaction was made slightly acidic (pH=5) with concentrated HCI (a few drops). The desired product was isolated directly by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.1 64 g, 44%. lon(s) found by LCMS: [(M-2Boc)+2H]/2 = 535.8, [(M-3Boc)+2H]/2 =485.8 Step c. Synthesis of Cbz-deca-Boc Intermediate

A solution of (2S,5S,8S,16S,19S,22S)-12-[(benzyloxy)carbonyl]-2,8,16,22-tetrakis{2-[(tert- butoxycarbonyl)amino]ethyl}-5,19-bis[(1 R)-1 -hydroxyethyl]-4,7,10,14,17,20-hexaoxo-3,6,9,12,1 5,1 8,21 - heptaazatricosane-1 ,23-dioic acid (0.164 g, 0.129 mmol), INT-1 1 (0.329 g, 0.310 mmol), DIEA (0.146 mL, 0.839 mmol), and DMF (1 mL), was treated with a solution of HATU (0.175 g, 0.460 mmol) in DMF (1 mL), added dropwise over 30 minutes. The crude reaction mixture was taken on to the next step without purification, lon(s) found by LCMS: [(M-2Boc)+3H]/3 = 1 053.3, [(M-3Boc)+3H]/3 = 1 019.9, [(M- 4Boc)+3H]/3 = 986.6. Step d. Synthesis of deca-Boc Intermediate

Crude Cbz-deca-Boc Intermediate from Step c (DMF solution) was diluted with methanol (10 mL), charged with 5%Pd/C (0.1 50 g), and hydrogen from a balloon. The reaction was monitored by LCMS. After 2 hr the mixture was filtered through Celite, concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.252 g, 61 % (two steps), lon(s) found by LCMS: (M+3H)/3 = 1 075.3

Step e. Synthesis of deca-Boc-(Compound 11)

A solution of deca-Boc-intermediate from Step d (0.100 g, 0.031 mmol), 4-[(2-{2-[(6-deoxy-alpha- L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid (INT-2) (0.012 g, 0.0341 mmol, and DIEA (0.016 mL, 0.093 mmol), in DMF (1 mL), were treated with HATU (0.014 g, 0.037 mmol), while stirring at room temperature. After 30 minutes, the product was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield 0.095 g, 86% yield, lon(s) found by LCMS: [(M-2Boc)+3H]/3 = 1 1 19.7, [(M- 3Boc)+3H]/3 = 1086.3

Step f. Synthesis of Compound 11

A solution of deca-Boc-(Compound 1 1 ) (0.095 g, 0.027 mmol), dissolved in DCM (1 mL), was treated with TFA (1 mL), while stirring at room temperature. After 5 minutes, the product was

concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 0.061 g, 75% yield, lon(s) found by LCMS: (M+3H)/3 = 852.8, (M+4H)/4 = 639.8 Example 34.

Compound 12 was prepared analogously to Compound 1 1 , where 4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid was substituted with 1 -[(6-deoxy-alpha-L- mannopyranosyl)oxy]-7,10-dioxo-3,14,17,20,23-pentaoxa-6,1 1 -diazahexacosan-26-oic acid in the first step of the sequence, lon(s) found by LCMS: (M+4H)/4 = 701 .6, (M+5H)/5 = 561 .5

Example 35. Synthesis of Compo

Step a. Synthesis of 5-(2-{[(benzyloxy)carbonyl]amino}ethoxy)benzene-1 ,3-dicarboxylic acid

Dimethyl-5-hydroxyisophthalate (1 .0 g, 4.76 mmol), benzyl (2-bromoethyl)carbamate (1 .5 g, 6.19 mmol) and cesium carbonate (1 .6 g, 4.76 mmol) were stirred together in DMF (3 mL) at 50 °C for 12 hours. The mixture was cooled, filtered and applied directly to reversed phase HPLC (20-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were concentrated and stirred in a 1 /1 /2 mixture of methanol/THF/DI water (15 mL) containing lithium hydroxide (0.68 g, 28.6 mmol) at room temperature for 2 hours. The mixture was neutralized with 1 N HCI aqueous, extracted into ethyl acetate, concentrated and applied to reversed phase HPLC (10-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and lyophilized to afford 5-(2-{[(benzyloxy)carbonyl]amino}ethoxy)benzene-1 ,3-dicarboxylic acid as a white solid. Yield: 79%, 2 steps. LC/MS [M-H-] - 358.4. Step b. Synthesis of (2S,2'S)-2,2'-([5-(2-{[(benzyloxy)carbonyl]amino}ethoxy)-1 ,3- phenylene]bis{carbonylazanediyl[(2S,3R)-3-hydroxy-1-oxobutane-2,1-diyl]azanediyl})bis{4-[(tert- butoxycarbonyl)amino]butanoic acid}

HATU (550 mg, 1 .22 mmol) was added to a stirring mixture of 5-(2- {[(benzyloxy)carbonyl]amino}ethoxy)benzene-1 ,3-dicarboxylic acid (200 mg, 0.56 mmol), INT-21 (408 mg, 0.122 mmol), DIEA (431 mg, 3.34 mmol) in DMF (4 mL). The mixture was stirred for 30 minutes and then applied directly to reversed phase HPLC (20-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were concentrated and stirred in a 1 /1 /2 mixture of methanol/THF/DI water (15 mL) containing lithium hydroxide (80 mg, 3.3 mmol) at room temperature for 2 hours. The mixture was neutralized with 1 N HCL aqueous and extracted into ethyl acetate, concentrated and applied to reversed phase HPLC (1 0-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and lyophilized to afford (2S,2'S)-2,2'-([5-(2- {[(benzyloxy)carbonyl]amino}ethoxy)-1 ,3-phenylene]bis{carbonylazanediyl[(2S,3R)-3-hydroxy-1 - oxobutane-2,1 -diyl]azanediyl})bis{4-[(tert-butoxycarbonyl)amino]butanoic acid} as a white solid. Yield: 88 %, 2 steps. LC/MS (M+2H)/2 = 481 .2.

Step c. Synthesis of hexa-tert-butyl ([5-(2-aminoethoxy)-1 ,3- phenylene]bis{carbonylazanediyl[(2S,3R)-3-hydroxy-1-oxobutane-2,1-diyl]azanediyl{(2S)-4-[(tert- butoxycarbonyl)amino]-1-oxobutane-2,1-diyl}azanediyl[(2S,5R,8S,11S,14S,17S,22S)-5-benzyl-17- [(1 R)-1-hydroxyethyl]-8-(2-methylpropyl)-3,6,9,12,15,18,23-heptaoxo-1 ,4,7,10,13,16,19- heptaazacyclotricosane-22,2,11 ,14-tetrayl]tri(ethane-2,1-diyl)})hexakiscarbamate

HATU (174 mg, 0.457 mmol) was added to a stirring mixture of (2S,2'S)-2,2'-([5-(2- {[(benzyloxy)carbonyl]amino}ethoxy)-1 ,3-phenylene]bis{carbonylazanediyl[(2S,3R)-3-hydroxy-1 - oxobutane-2,1 -diyl]azanediyl})bis{4-[(tert-butoxycarbonyl)amino]butanoic acid} (200 mg, 0.208 mmol), INT-1 1 (485 mg, 457 mmol) and DIEA (161 mg, 1 .25 mmol) in DMF (4 mL). The mixture was stirred for 30 minutes and then applied directly to reversed phase HPLC (30-95% methanol in Dl water containing 0.1 % formic acid: 25 minute gradient). The pure fractions were pooled and concentrated. The CBZ-protected intermediate was taken up in methanol (15 m) and stirred for 1 hour in the presence of 5% Pd/C (75 mg) under 1 atmosphere of hydrogen. The mixture was filtered, concentrated and applied to reversed phase HPLC (20-95% methanol in Dl water containing 0.1 % formic acid: 25 minute gradient). The pure fractions were pooled and concentrated to afford hexa-tert-butyl ([5-(2-aminoethoxy)-1 ,3- phenylene]bis{carbonylazanediyl[(2S,3R)-3-hydroxy-1 -oxobutane-2,1 -diyl]azanediyl{(2S)-4-[(tert- butoxycarbonyl)amino]-1 -oxobutane-2,1 -diyl}azanediyl[(2S,5R,8S,1 1 S,14S,17S,22S)-5-benzyl-17-[(1 R)- 1 -hydroxyethyl]-8-(2-methylpropyl)-3,6,9,12,15,18,23-heptaoxo-1 ,4,7,1 0,13,16,19- heptaazacyclotricosane-22,2,1 1 ,14-tetrayl]tri(ethane-2,1 -diyl)})hexakiscarbamate as a white solid. Yield: 66 %, 2 steps. LC/MS [m-boc/3+H+] = 872.5.

Step d. Synthesis of Compound 13

HATU was added to a stirring mixture of 4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid (1 8 mg, 0.051 mmol) , hexa-tert-butyl ([5-(2- aminoethoxy)-1 ,3-phenylene]bis{carbonylazanediyl[(2S,3R)-3-hydroxy-1 -oxobutane-2,1 - diyl]azanediyl{(2S)-4-[(tert-butoxycarbonyl)amino]-1 -oxobutane-2,1 - diyl}azanediyl[(2S,5R,8S ,1 1 S ,14S ,17S,22S)-5-benzyl-17-[(1 R)-1 -hydroxyethyl]-8-(2-methylpropyl)- 3,6,9,12,15,18,23-heptaoxo-1 ,4,7,10,13,16,19-heptaazacyclotricosane-22,2,1 1 ,14-tetrayl]tri(ethane-2,1 - diyl)})hexakiscarbamate (100 mg, 0.034 mmoL) and DIEA (13 mg, 0.103 mmol) in DMF (1 .5 mL). The mixture was stirred for 30 minutes and then applied directly to reversed phase HPLC (30-95% acetonitrile in Dl water containing 0.1 % formic acid: 25 minute gradient). The pure fractions were pooled and concentrated and the resulting white solid was stirred in a 1 /1 mixture of TFA/DCM containing 0.1 % thioanisole (4 mL) at room temperature for 30 minutes. The solvent was removed by rotovap, azeotroped twice with toluene. The residue was dissolved in Dl water (1 mL) and applied to reversed phase HPLC (0- 35% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were pooled and lyophilized to afford the title compound (Compound 13) as a white solid. Yield: 23%, 2 steps. LC/MS (M+3H)/3 = 816.8. Example 36. Synthesis of Compound 14

Step a. Synthesis of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-

{[(benzyloxy)carbonyl]amino}octanoyl]amino}-4-[(tert-butoxycarbonyl)amino]butanoyl}-L- threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate

A stirring solution of methyl (2S)-2-[(N-{(2S)-2-amino-4-[(tert-butoxycarbonyl)amino]butanoyl}-L- threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.300 g, 0.562 mmol), (2S)-2- {[(benzyloxy)carbonyl]amino}octanoic acid (0.165 g, 0.562 mmol), and DIEA (0.294 mL, 1 .69 mmol), in DMF (1 mL), were treated with HATU (0.235 g, 0.618 mmol) at room temperature. The desired product was isolated by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using 0.1 % TFA as the modifier. Yield 0.296g, 65%. lon(s) found by LCMS: (M+H)+ = 808.5 Step b. Synthesis of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-aminooctanoyl]amino}-4-[(tert- butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate

A solution of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-{[(benzyloxy)carbonyl]amino}octanoyl]amino}-4- [(tert-butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (1 .160 g, 1 .433 mmol), in methanol (5 mL), was charged with 5%Pd/C (0.400 g), and hydrogen from a balloon. When the reaction was complete by LCMS (1 hr), the mixture was filtered through Celite, concentrated and taken-on to the next step without purification. Yield 0.630 g, 65%. lon(s) found by LCMS: (M+H)+ = 675.4

Step c. Synthesis of dimethyl (2S,5S,8S,11S,19S,22S,25S,28S)-15-[(benzyloxy)carbonyl]-2,8,22,28- tetrakis{2-[(tert-butoxycarbonyl)amino]ethyl}-11 ,19-dihexyl-5,25-bis[(1 R)-1-hydroxyethyl]- 4,7,10,13,17,20,23,26-octaoxo-3,6,9,12,15,18,21 , 24,27-nonaazanonacosane-1 ,29-dioate

A solution of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-aminooctanoyl]amino}-4-[(tert- butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.626 g, 0.928 mmol), 2,2'-{[(benzyloxy)carbonyl]azanediyl}diacetic acid (0.124 g, 0.464 mmol), and DIEA(0.364 mL, 2.088 mmol) in DMF (3 mL), were treated with a solution of HATU (0.362 g, 0.951 mmol) in DMF (1 mL), dropwise over 15 minutes. The desired product was isolated by reversed phase liquid

chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.383 g, 52%. lon(s) found by LCMS: [(M-2Boc)+2H]/2 =690.9, [(M-4Boc)+2H]/2 = 590.9

Step d. Synthesis of (2S,5S,8S,11S,19S,22S,25S,28S)-15-[(benzyloxy)carbonyl]-2,8,22,28-tetrakis{2-

[(tert-butoxycarbonyl)amino]ethyl}-11 ,19-dihexyl-5,25-bis[(1 R)-1-hydroxyethyl]-

4,7,10,13,17,20,23,26-octaoxo-3,6,9,12,15,18,21 , 24,27-nonaazanonacosane-1 ,29-dioic acid

A solution of dimethyl (2S,5S,8S,1 1 S,19S,22S,25S,28S)-15-[(benzyloxy)carbonyl]-2, 8,22,28- tetrakis{2-[(tert-butoxycarbonyl)amino]ethyl}-1 1 ,19-dihexyl-5,25-bis[(1 R)-1 -hydroxyethyl]-

4,7,10,13,17,20,23,26-octaoxo-3,6,9,12,15,18,21 ,24,27-nonaazanonacosane-1 ,29-dioate (0.383 g, 0.242 mmol), in hot methanol (10 mL) was treated with a solution of lithium hydroxide (0.0232 g, 0.969 mmol), in water (2 mL), while stirring at room temperature. The desired product was isolated by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield 0.340 g, 90%. lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 726.9, [(M-2Boc)+2H]/2 = 676.9

Step e. Synthesis of Cbz-deca-Boc-(Compound 14) Precursor

A solution of (2S,5S,8S,1 1 S,19S,22S,25S,28S)-15-[(benzyloxy)carbonyl]-2,8,22,28-tetrakis{2- [(tert-butoxycarbonyl)amino]ethyl}-1 1 ,19-dihexyl-5,25-bis[(1 R)-1 -hydroxyethyl]-4,7,10,13,17,20,23,26- octaoxo-3,6,9,12,1 5,1 8,21 ,24,27-nonaazanonacosane-1 ,29-dioic acid (0.357 g, 0.230 mmol), INT-1 1 (0.488 g, 0.460 mmol), DIEA (0.191 mL, 1 .095 mmol), and DMF (5 mL), was treated with HATU (0.175 g, 0.460 mmol). The crude reaction mixture was taken on to the next step without purification, lon(s) found by LCMS: [(M-3Boc)+3H]/3 = 1 1 14.0

Step f. Synthesis of deca-Boc-(Compound 14) Precursor

Crude Cbz-deca-Boc-(Compound 14) Precursor was diluted with methanol (1 0 mL), charged with 5%Pd/C (0.250 g), and hydrogen from a balloon. The reaction was monitored by LCMS. After 2 hr the mixture was filtered through Celite, concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield 0.139 g, 18% (two steps), lon(s) found by LCMS: [(M-2Boc)+3H]/3 = 1 102.6

Step g. Synthesis of deca-Boc-(Compound 14)

Deca-Boc-(Compound 14) Precursor (0.070 g, 0.0200 mmol), INT-2 (0.042 g, 0.120 mmol), DIEA (0.063 mL, 0.359 mmol), and DMF (1 mL) were treated with HATU (0.046 g, 0.120 mmol), while stirring at room temperature. The reaction was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.070 g, 91 % yield, lon(s) found by LCMS: [(M-2Boc)+3H]/3 = 1213.7, [(M-3Boc)+3H]/3 = 1 1 80.4, [(M-4Boc)+3H]/3 = 1 147.0

Step h. Synthesis of Compound 14

Deca-Boc-(Compound 14) (0.070 g, 0.01 82 mmol) was dissolved in DCM (2 mL) and treated with TFA (1 mL ), while stirring at room temperature. After 30minutes, the reaction was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid(0.1 %) as the modifier. Yield of Compound 14 was 0.013g, 25% yield, lon(s) found by LCMS: (M+4H)/4 = 710.4, (M+5H)/5 = 568.5, (M+6H)/6 = 473.9

Step a. Synthesis of Cbz-deca-Boc-(Compound 14) Precursor

A solution of INT-20 (2.23 g, 1 .308 mmol), 2,2'-{[(benzyloxy)carbonyl]azanediyl}diacetic acid

(0.175 g, 0.654 mmol), DIEA (0.718 mL, 4.12 mmol), and DMF (10 mL), was treated with a solution HATU (0.522 g, 1 .373 mmol) in DMF (3 mL) via syringe pump over 1 hr, while stirring at room temperature. The crude reaction mixture was taken on to the next step without purification, lon(s) found by LCMS: [(M- 3Boc)+3H]/3 = 1 1 14.0

Step b. Synthesis of deca-Boc-(Compound 14) Precursor

Crude Cbz-deca-Boc-(Compound 14) Precursor in DMF, was charged with 5% Pd/C (1 .0 g), and hydrogen gas from a balloon. The reaction was monitored by LCMS. After 2 hr the mixture was filtered through Celite, concentrated and purified by C18 reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 20% to 100% methanol and water, using no modifier. Yield 1 .46 g, 64% (two steps), lon(s) found by LCMS: [(M-2Boc)+3H]/3 = 1 1 02.6

Step c. Synthesis of deca-Boc-(Compound 14)

A solution of deca-Boc-(Compound 14) Precursor (1 .46 g, 0.41 6 mmol), 4-[(2-{2-[(6-deoxy-alpha- L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid (INT-2) (0.161 g, 0.458 mmol), DIEA

(0.239 mL, 1 .374 mmol), and DMF (5 mL) were treated with a solution of HATU (0.174 g, 0.457 mmol) in DMF (3 mL), dropwise over 30 minutes, while stirring at room temperature. The reaction was purified by C18 reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 30% to 100% methanol and water, using no modifier. Yield 1 .56 g, 97% yield, lon(s) found by LCMS: [(M-2Boc)+3H]/3 = 1213.7, [(M-3Boc)+3H]/3 = 1 180.4, [(M-4Boc)+3H]/3 = 1 147.0 Step d. Synthesis of Compound 14

Deca-Boc-(Compound 14) (1 .56 g, 0.406 mmol) was dissolved in DCM (10 mL) and treated with TFA (10 mL), while stirring at room temperature for 5 minutes. The reaction was concentrated at 0°C via rotary evaporation, stored under high vacuum for 1 hr to remove residual TFA, dissolved in methanol and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 0% to 30% acetonitrile and water over 20 minutes, using formic acid (0.1 %) as the modifier. After analysis by HPLC the desired fractions are pooled and concentrated to colorless foam. Yield of Compound 14 was 1 .18 g, 96% yield, lon(s) found by LCMS: (M+4H)/4 = 710.4, (M+5H)/5 = 568.5, (M+6H)/6 = 473.9

E

Step a. Synthesis of Cbz-deca-Boc-(Compound 15) Precursor

A stirring solution of INT-17 (0.200 g, 0.131 mmol), Cbz-L-aspartic acid (0.017 g, 0.065 mmol), and DIEA (0.068 mL, 0.392 mmol), dissolved in DMF (1 mL), was treated with a solution of HATU (0.050 g, 0.0131 mmol) in DMF (1 .0 mL), dropwise over 30 minutes. After 1 hour the product was taken on to the next step without further purification, lon(s) found by LCMS: [(M-3Boc)+3H]/3 = 997.3, [(M- 4Boc)+3H]/3 = 964.0, [(M-5Boc)+3H]/3 = 930.6 Step b. Synthesis of deca-Boc-(Compound 15) Precursor

A solution of Cbz-deca-Boc-(Compound 15) Precursor (431 mg, 0.131 mmol) dissolved in methanol (5 mL), was charged with 5% Pd/C (0.050 g), flushed with hydrogen from a balloon, and stirred under a hydrogen atmosphere until Cbz removal was complete by LCMS (~0.5hr). The crude product was isolated by filtration through Celite, then concentrated to an oil, which was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield for two steps 0.147 g, 71 % yield, lon(s) found by LCMS: [(M-3Boc)+3H]/3 = 952.6, [(M-4Boc)+3H]/3 = 91 9.3 Step c. Synthesis of deca-Boc-Compound 15

A solution of deca-Boc-(Compound 15) Precursor (0.1 06 g, 0.0335 mmol), INT-2 (0.013 g, 0.0370 mmol), and DIEA (0.019 mL, 0.1 1 1 mmol), in DMF (1 mL), were treated with HATU (0.046 g, 0.120 mmol), while stirring at room temperature. After 30 minutes, the product was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield 0.1 15 g, 98% yield, lon(s) found by LCMS: [(M- 2Boc)+3H]/3 = 1097.0, [(M-3Boc)+3H]/3 = 1063.7

Step d. Synthesis of Compound 15

A solution of deca-Boc-(Compound 15) (0.109 g, 0.031 mmol) in DCM (1 mL) was treated TFA (1 mL), while stirring at room temperature. After 5 minutes the solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% acetonitrile and water, using formic acid as the modifier. Yield of Compound 15 was 0.033 g, 40% yield, lon(s) found by LCMS: (M+3H)/3 = 830.2, (M+4H)/4 =622.9, (M+5H)/5 =498.5. Example 39. Sy

A stirring solution of INT-17 (0.150 g, 0.098 mmol), fumaric acid (0.0057 g, 0.049 mmol), and DIEA (0.051 mL, 0.294 mmol) in DMF (1 mL), was treated with a solution of HATU (0.037 g, 0.098 mmol) in DMF (0.5 mL), dropwise over 15 minutes. After 1 hour the product was isolated by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield 0.108 g, 70% yield, lon(s) found by LCMS: [(M- 3Boc)+3H]/3 = 947.0, [(M-4Boc)+3H]/3 = 913.6, [(M-5Boc)+3H]/3 = 880.3

Step a. Synthesis of Benzyl-deca-Boc-(Compound 16) Precursor

Benzyl-deca-Boc-(Compound 1 6) Precursor was prepared analogously as in Example 39, where 1 -benzyl-3,3-azetidinedicarboxylic acid, was substituted for fumaric acid, lon(s) found by LCMS: [(M- 2Boc)+3H]/3 = 1020.0, [(M-3Boc)+3H]/3 = 986.6

Step b. Synthesis of deca-Boc-(Compound 16) Precursor

A solution of benzyl-deca-Boc-(Compound 16) Precursor (0.175 g, 0.537 mmol) dissolved in methanol (5 mL), was charged with 5% Pd/C (0.050 g), flushed with hydrogen gas from a balloon, and stirred under a hydrogen atmosphere until debenzylation was complete by LCMS (~2hr). The crude product was isolated by filtration through Celite, then concentrated to an oil, and used in the next step without further purification. Step c. Synthesis of deca-Boc-(Compound 16)

A solution of deca-Boc-(Compound 16) Precursor (0.175 g, 0.0552 mmo), 4-[(2-{2-[(6-deoxy- alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid (INT-2) (0.0291 g, 0.0828 mmol) and DIEA (0.029 mL, 0.166 mmol), in DMF(1 mL ), was treated with a solution of HATU (0.0220 g, 0.0580 mmol) in DMF (0.5 mL ), dropwise over 30 minutes. After stirring for 1 hr at room temperature, the product was isolated by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.089 g, 46%. lon(s) found by LCMS: [(M-1 Boc)+3H]/3 = 1 134.3, [(M-2Boc)+3H]/3 = 1 1 01 .0 Step d. Synthesis of Compound 16

A solution of deca-Boc-(Compound 16) (0.025 g, 0.00767 mmol), dissolved in methylene chloride (1 mL) was treated with TFA (1 mL), while stirring at room temperature. After 5 minutes, the product was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 0.01 Og , 58% yield, lon(s) found by LCMS: (M+2H)/2 = 1250.7, (M+3H)/3 = 834.2

Example 41. Synthesis of Compound 17

Step a. Synthesis of the dimethyl ester linker

To a solution of a mixture of 14-{2-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}-4,12,16,24-tetraoxo-8,20-dioxa-5,1 1 ,14,17,23- pentaazaheptacosane-1 ,27-dioic acid (from Example 23, Step f) (24.9 mg, 0.0312 mmol), methyl (2S)-2- [(N-{(2S)-2-amino-4-[(tert-butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert- butoxycarbonyl)amino]butanoate (36.8 mg, 0.07 mmol), and EteN (20.2 mg, 0.2 mmol) in anhydrous DMF (0.5 mL) was added dropwise a solution of HATU (26.2 mg, 0.069 mmol) in anhydrous DMF (0.5 mL) over 30 minutes. The reaction was stirred for 1 hour, then directly purified through RPLC (15 to 55 % acetonitrile and water). Yield 48.7 mg, 85.4%. lon(s) found by LCMS: [(M - Boc)/2]+1 = 864.5.

Step b. Synthesis of the linker

The dimethyl ester linker (Step a) was dissolved in MeOH (1 mL). After the solution was cooled in an ice-water bath, a solution of LiOH (10 mg, 0.25 mmol) was added. The reaction was stirred for 2 hours, then acidified by formic acid (0.02 mL) and purified through RPLC (5 to 78 % MeOH and water). Yield 46.4 mg, 96.9%. lon(s) found by LCMS: [(M - Boc)/2]+1 = 850.

Step c. Synthesis of Compound 17

To a solution of a mixture of the linker (Step b) (46.4 mg, 0.026 mmol), INT-1 1 (60.6 mg, 0.057 mmol), and EteN (1 0.1 mg, 0.1 mmol) in anhydrous DMF (1 mL) was added dropwise a solution of HATU (21 mg, 0.055 mmol) in anhydrous DMF (0.5 mL) over 30 minutes. The reaction was stirred for 1 hour, then directly purified through RPLC (25 to 100 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (79.5 mg, LCMS: [(M - Boc)/3]+1 = 872). The material was dissolved in TFA (~ 0.5 mL). After the solution was stirred for 15 minutes, it was purified by HPLC (3 to 12 % acetonitrile and water). Yield 10.8 mg, 10.3%. lon(s) found by LCMS: (M+4H)/4 = 722.

Example 42. Synth

The title compound was prepared analogously to Compound 1 but utilized 2,2'-({2-[(2-{2-[(6- deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)diacetic acid (Example 23, Step d) as the linker, lon(s) found by LCMS (M+2H)/2 = 1 157.6, (M+3H)/3 =772.1 , (M+4H)/4 =579.3. (M+5H)/5 =463.7. Example 43. Synthesis of Compound 19

Step a. Synthesis of dimethyl 5-(2-aminoethoxy)benzene-1 ,3-dicarboxylate

Dimethyl-5-hydroxyisophthalate (2 g, 9.5 mmol), benzyl (2-bromoethyl)carbamate (1 .6 g, 12.4 mmol) and cesium carbonate (3 g, 9.5 mmol) were stirred together in DMF (6 mL) at 50 °C for 12 hours. The mixture was cooled, filtered and applied directly to reversed phase HPLC (20-95% acetonitrile in Dl water containing 0.1 % formic acid: 20 minute gradient). The pure fractions were concentrated and stirred in a mixture of methanol (25 mL) and 5% Pd/C (200 mg) under 1 atmosphere of hydrogen gas for 12 hours. The mixture was filtered, concentrated and purified by reversed phase HPLC (5-95% acetonitrile in Dl water with no modifier: 20 minute gradient) to afford dimethyl 5-(2-aminoethoxy)benzene-1 ,3- dicarboxylate as a clear oil. Yield: 84%, 2 steps. LC/MS [M+H+]⁺ 254.2. Step b. Synthesis of 5-{2-[4-(1-{2-[2-(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethoxy)ethoxy]ethyl}-1 H-1 ,2,3-triazol-4- yl)butanamido]ethoxy}benzene-1 ,3-dicarboxylic acid

HATU (419 mg, 1 .1 mmol, in 1 mL DMF) was added to a stirring mixture of 4-(1 -{2-[2-(2-{2-[(6- deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethoxy)ethoxy]ethyl}-1 H-1 ,2,3-triazol-4-yl)butanoic acid (526 mg, 1 .1 mmol), dimethyl 5-(2-aminoethoxy)benzene-1 ,3-dicarboxylate (254 mg, 1 .0 mmol) and diisopropylethylamine (259 mg, 2.0 mmol) in DMF (3 mL). The mixture was stirred for 45 minutes then applied directly to reversed phase HPLC (5-95% acetonitrile in Dl water containing 0.1 % TFA: 20 minute gradient). The pure fractions were concentrated and stirred in 1 /1 /2 mixture of THF/methanol/DI water (15 mL) containing lithium hydroxide (144 mg, 6.0 mmol) for 20 minutes. The mixture was acidified with TFA (~1 mL) and the volume was concentrated by half on the rotary evaporator. The mixture was applied to reversed phase HPLC (0-40% acetonitrile in Dl water containing 0.1 % TFA:20 minute gradient). The pure fractions were pooled and lyophilized to afford 5-{2-[4-(1 -{2-[2-(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethoxy)ethoxy]ethyl}-1 H-1 ,2,3-triazol-4-yl)butanamido]ethoxy}benzene-1 ,3- dicarboxylic acid as a light greenish colored oil. Yield: 29%, 2 steps. LC/MS [m/2-H]- 683.4.

Step c. Synthesis of (2R)-2-[3-{[(1S)-1-carboxy-2-(naphthalen-2-yl)ethyl]carbamoyl}-5-(2-{4-[1-(2- {2-[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}ethoxy)ethoxy]ethoxy}ethyl)- 1 H-1 ,2,3-triazol-4-yl]butanamido}ethoxy)benzamido]-3-(naphthalen-2-yl)propanoic acid

HATU (347 mg, 0.92 mmol, in 1 mL DMF) was added to a stirring mixture of 5-{2-[4-(1 -{2-[2-(2-{2-

[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethoxy)ethoxy]ethyl}-1 H-1 ,2,3-triazol-4- yl)butanamido]ethoxy}benzene-1 ,3-dicarboxylic acid (285 mg, 0.42 mmol), methyl (2R)-2-amino-3- (naphthalen-2-yl)propanoate hydrochloride (243 mg, 0.92 mmol) and triethylamine (252 mg, 2.5 mmol) in DMF (2 mL). The mixture was stirred for 45 minutes then applied directly to reversed phase HPLC (5-95% acetonitrile in Dl water containing 0.1 % TFA: 20 minute gradient). The pure fractions were concentrated and stirred in 1 /1 /2 mixture of THF/methanol/DI water (10 mL) containing lithium hydroxide (60 mg, 2.5 mmol) for 20 minutes. The mixture was acidified with TFA (~1 mL) and the volume was concentrated by half on the rotary evaporator. The mixture was applied to reversed phase HPLC (0-40% acetonitrile in Dl water containing 0.1 % TFA: 20 minute gradient). The pure fractions were pooled and lyophilized to afford (2R)-2-[3-{[(1 S)-1 -carboxy-2-(naphthalen-2-yl)ethyl]carbamoyl}-5-(2-{4-[1 -(2-{2-[2-(2-{[(2R,3R,4R,5R,6S)- 3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}ethoxy)ethoxy]ethoxy}ethyl)-1 H-1 ,2,3-triazol-4- yl]butanamido}ethoxy)benzamido]-3-(naphthalen-2-yl)propanoic acid as a light greenish colored oil. Yield: 43%, 2 steps. LC/MS [M+H⁺]⁺ 1079.4. Step d. Synthesis of N~1 ~,N~3~-bis[(2S)-1-{[(2S,3R)-1-{[(2S)-4-amino-1-oxo-1-

{[(3S,6S,9S,12S,15R,18S,21S)-6,9,18-tris(2-aminoethyl)-15-benzyl-3-[(1 R)-1-hydroxyethyl]-12-(2- methylpropyl)-2,5,8,11 ,14,17,20-heptaoxo-1 , 4,7,10,13, 16,19-heptaazacyclotricosan-21 - yl]amino}butan-2-yl]amino}-3-hydroxy-1-oxobutan-2-yl]amino}-3-(naphthalen-2-yl)-1-oxopropan-2- yl]-5-(2-{4-[1-(2-{2-[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2- yl]oxy}ethoxy)ethoxy]ethoxy}ethyl)-1 H-1 ,2,3-triazol-4-yl]butanamido}ethoxy)benzene-1 ,3- dicarboxamide (Compou

HATU (50 mg, 0.13 mmol, in 0.5 mL DMF) was added dropwise over 30 minutes to a stirring mixture of INT-16 (181 mg, 0.13 mmol), (2R)-2-[3-{[(1 S)-1 -carboxy-2-(naphthalen-2-yl)ethyl]carbamoyl}-5- (2-{4-[1 -(2-{2-[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2- yl]oxy}ethoxy)ethoxy]ethoxy}ethyl)-1 H-1 ,2,3-triazol-4-yl]butanamido}ethoxy)benzamido]-3-(naphthalen-2- yl)propanoic acid (65 mg, 0.060 mmol), and triethylamine (36 mg, 0.36 mmol) in DMF (1 .5 mL) and the reaction was stirred for an additional 30 minutes at room temperature. The mixture was applied directly to reversed phase HPLC (20-95% methanol in Dl water containing no modifier: 20 minute gradient). The pure fractions containing the desired intermediate (LC/MS (M+3H)/3⁺1 156.8:loss of 2 Bocs) were pooled, concentrated and dried under high vacuum. The Boc-protected intermediate was stirred in a 1 /1 mixture of TFA/DCM containing thioanisole (60 mg, 0.482 mmol) for 20 minutes at ambient temperature. The solvent was removed on the rotary evaporator and purified by to reversed phase HPLC (0-35% acetonitrile in Dl water containing 0.1 % TFA: 20 minute gradient). The pure fractions were pooled and lyophilized to afford N~1 ~,N~3~-bis[(2S)-1 -{[(2S,3R)-1 -{[(2S)-4-amino-1 -oxo-1 - {[(3S,6S,9S,12S,15R,18S.21 S)-6,9,18-tris(2-aminoethyl)-15-benzyl-3-[(1 R)-1 -hydroxyethyl]-12-(2- methylpropyl)-2,5,8,1 1 ,14,17,20-heptaoxo-1 ,4,7,1 0,13,16,1 9-heptaazacyclotricosan-21 -yl]amino}butan-2- yl]amino}-3-hydroxy-1 -oxobutan-2-yl]amino}-3-(naphthalen-2-yl)-1 -oxopropan-2-yl]-5-(2-{4-[1 -(2-{2-[2-(2- {[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}ethoxy)ethoxy]ethoxy}ethyl)-1 H-1 ,2,3-triazol- 4-yl]butanamido}ethoxy)benzene-1 ,3-dicarboxamide (Compound 19) as a white solid. Yield: 41 %, 2 steps. LC/MS (M+3H)/3⁺ 990.3.

Step a. Synthesis of di-tert-butyl 37-(2-(benzyloxy)-2-oxoethyl)-35,39-dioxo- 4,7,10,13,16,19,22,25,28,31 , 43,46,49,52,55,58,61 ,64,67,70-icosaoxa-34,37,40-triazatriheptacontane- 1 ,73-dioate

To the solution of 2,2'-((2-(benzyloxy)-2-oxoethyl)azanediyl)diacetic acid (70 mg, 0.25 mmol) and amino-PEG10-tert-butyl ester (290 mg, 0.5 mmol) in DMF (5 ml_) was added EDC (120 mg, 0.6 mmol), HOBT (90 mg, 0.6 mmol) and Hunig's base (1 .4 ml_, 10 mmol) at room temperature. The solution was stirred overnight. The reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, 21 0 mg 75% yield. LC/MS (M+2H)/2 709.0.

Step b. Synthesis of 3-(38,38-dimethyl-2,36-dioxo-6,9,12,15,18,21 , 24,27,30,33,37-undecaoxa-3- azanonatriacontyl)-41 ,41-dimethyl-5,39-dioxo-9,12,15,18,21 ,24,27,30,33,36,40-undecaoxa-3,6- diazadotetracontan-1-oic acid

Di-tert-butyl 37-(2-(benzyloxy)-2-oxoethyl)-35,39-dioxo- 4,7,10,13,16,19,22,25,28,31 ,43,46,49,52,55,58,61 ,64,67,70-icosaoxa-34,37,40-triazatriheptacontane- 1 ,73-dioate (210 mg, 0.15 mmol) was dissolved into a mixture of 5 mL MeOH and 5 mL ethyl acetate, then 0.1 g of 5% palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere overnight. The palladium on charcoal was removed by filtration after completion of the reaction as determined by LCMS. The filtrate was concentrated and used in the next step without any purification. LC/MS [M/2-t-Butyl]+1 663.9. Step c. Synthesis of 35,39-dioxo-37-(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yl)oxy)ethoxy)ethyl)amino)ethyl)-

4,7,10,13,16,19,22,25,28,31 , 43,46,49,52,55,58,61 ,64,67,70-icosaoxa-34,37,40-triazatriheptacontane- 1 ,73-dioic acid

To a solution of 3-(38,38-dimethyl-2,36-dioxo-6,9,12,15,18,21 ,24,27,30,33,37-undecaoxa-3- azanonatriacontyl)-41 ,41 -dimethyl-5,39-dioxo-9,12,1 5,18,21 ,24,27,30,33,36,40-undecaoxa-3,6- diazadotetracontan-1 -oic acid (0.1 5 mmol) and INT-1 (50 mg, 0.2 mmol) in DMF (5 mL) was added EDC (20 mg, 0.1 mmol), HOBT (15mg, 0.1 mmol) and Hunig's base (0.3 mL, 0.2 mmol) at room temperature. The solution was stirred overnight. The reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil, 150 mg 75% yield. LC/MS [M/2-2X t-Butyl]+1 724.4.

The above product was treated with TFA and stirred for 30 minutes, concentrated and dried under high vacuum and used in the next step without purification.

Step d. Prepa

To a solution of 14-{2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}-4,12,16,24-tetraoxo-8,20-dioxa-5,1 1 ,14,17,23-pentaazaheptacosane-1 ,27-dioic acid (150 mg, 0.1 mmol), triethylamine (0.07 mL, 0.5 mmol) and INT-16 (270 mg, 0.2 mmol) in 5 mL DMF was added HATU (78 mg, 0.2 mmol). The reaction was stirred for 1 hr and then the reaction mixture was

concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of product, 1 00 mg, 35% yield.

Per-Boc-(Compound 20) from the previous step was treated in 2 mL DCM and 2 mL TFA at room temperature, the solution was stirred 10 min, then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 0.015 g, 34% yield, lon(s) found by LCMS: (M+3H)/3 = 1 1 13.0, (M+4H)/4 = 835, (M+5H)/5 =668.2 (M+6H)/6=557. Example 45. Synth

To the solution of 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}azanediyl)diacetic acid (Example 23, Step d) (10 mg, 0.023 mmol), triethylamine (0.07 mL, 0.5 mmol) and INT-18 (74 mg, 0.047 mmol) in 5 mL DMF was added HATU (19 mg, 0.02 mmol) in 1 mL DMF by syringe pump over 1 hour. The reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of product, 30 mg, 37% yield.

The per-Boc-(Compound 21 ) from the previous step was treated in 2 mL DCM and 2 mL TFA at room temperature, the solution was stirred 10 min, then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield of Compound 21 was 0.01 5 g, 34% yield. lon(s) found by LCMS: (M+2H)/2 = 1257.7, (M+3H)/3 = 838.8, (M+4H)/4 =629.4, (M+5H)/5=503.7.

Example 46. Synthesis of (2S)-4-amino-2-((S)-6-(2-(((2S,3R)-1-(((S)-4-amino-1-oxo-1-

(((3S,6S,9S,12S,15R,18S,21S)-6,9,18-tris(2-aminoethyl)-15-benzyl-3-((R)-1-hydroxyethyl)-12- isobutyl-2,5,8,11 ,14,17,20-heptaoxo-1 , 4,7,10,13, 16,19-heptaazacyclotricosan-21 -yl)amino)butan-2- yl)amino)-3-hydroxy-1-oxobutan-2-yl)amino)-2-oxoethyl)-2-((R)-1-hydroxyethyl)-4,8,11-trioxo-9-(2- oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)ethoxy)ethyl)amino)ethyl)-17-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yl)oxy)-15-oxa-3,6,9,12-tetraazaheptadecanamido)-N-((3S,6S,9S,15R,18S,21 S)-6,9,18-tris(2-

aminoethyl)-12-benzyl-3-((R)-1-hydroxyethyl)-15-isobutyl-2,5,8,11 ,14,17,20-heptaoxo- 1 ,4,7,10,13,16,19

The title compound was prepared analogously to Compound 1 utilizing 6-(carboxymethyl)-4,8,1 1 - trioxo-9-(2-oxo-2-((2-(2-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)ethoxy)ethyl)amino)ethyl)-17-(((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yl)oxy)-15-oxa-3,6,9,12-tetraazaheptadecan-1 -oic acid (Example 28, Step d) as the linker, lon(s) found by LCMS: (M+2H)/2 = 1331 .7, (M+3H)/3 = 888.1 , (M+4H)/4 =666.4, (M+5H)/5 =533.3.

Step a. Synthesis of Cbz-D-Thr-Dab(Boc)-OH: (2S)-2-({N-[(benzyloxy)carbonyl]-D- threonyl}amino)-4-[(tert-butoxycarbonyl)amino]butanoic acid

Cbz-D-Thr-OH (commercial) (1 .92 g, 7.37 mmol), Dab(Boc)-OMe (commercial) (2.00 g, 7.37 mmol), EDCI (4.263 g, 1 1 .05 mmol), HOBt (1 .493 g, 1 1 .05 mmol) and NaHCOs (1 .238 g, 14.74 mmol) were weighed into a 100 mL round bottom flask. 50 mL of DCM/DMF (4:1 ) was added into the flask. The mixture was stirred at room temperature overnight (TLC or LC/MS monitoring). After completion, EtOAc (200 mL) was added to dilute the reaction mixture and washed with 1 N aq. HCI, saturated NaHCCb and brine. The organic layer was dried with Na2S04 and concentrated. The residue was purified with normal phase silica (2-7% MeOH/DCM) to give 3.00 g pure desired product as a white solid (87%). Ions were found as: positive charge (M-Boc+H⁺: 368.2), negative charge (M-H-: 467.2). The solid was dissolved in 10 mL of THF/water (1 :1 ) and treated with LiOH (0.95 equiv.) for 30 minutes at room temperature. The reaction mixture was acidified to pH~2 with 1 N aqueous HCI. THF was removed under reduced pressure. The residue was extracted with EtOAc. After drying and concentration, Cbz-D-Thr-Dab(Boc)-OH was obtained in quantitative yield. Ions were found as: positive charge (M-Boc+H⁺: 354.2), negative charge (M-H-: 452.2). Step b. Synthesis of D-Thr-Dab(Boc)-OMe: Methyl (2S)-2-({N-[(benzyloxy)carbonyl]-D- threonyl}amino)-4-[(tert-butoxycarbonyl)amino]butanoate~methyl (2S)-4-[(tert- butoxycarbonyl)amino]-2-(D-threonylamino)butanoate

(D-Thr-Dab(Boc)-OMe ) was obtained by hydrogenolysis of the compound from Step a. in quantitative yield. Ion was found as (M+H⁺: 334.2).

Step c. Synthesis of Cbz-Dab(Boc)-D-Thr-Dab(Boc)-OH : (2S)-2-[(N-{(2S)-2-

{[(benzyloxy)carbonyl]amino}-4-[(tert-butoxycarbonyl)amino]butanoyl}-D-threonyl)amino]-4-[(tert- butoxycarbonyl)amino]butanoic acid

D-Thr-Dab(Boc)-OMe (0.713 g, 2.14 mmol), Z-NH-Dab(Boc)-OH (DCHA salt) (1 .199 g, 2.25 mmol), EDCI (0.615 g, 3.21 mmol), HOBt (0.433 g, 3.21 mmol) and NaHCOs (0.359 g, 4.28 mmol) were weighed into a 100- mL round bottom flask. 20 mL of DCM/DMF (4:1 ) was added into the flask. The mixture was stirred at room temperature overnight. After completion, EtOAc (100 mL) was added to dilute the reaction mixture and washed with 1 N aq. HCI, saturated NaHCOe and brine. Dried with Na2S04 and condensed. The residue was purified with normal phase silica (2-7% MeOH/DCM) to give 1 .186 g pure desired product Cbz-Dab(Boc)-D-Thr-Dab(Boc)-OMe as a white solid (83%). Ion was found as (M-

Boc+H⁺: 568.2). This white solid was dissolved in 10 mL of THF/water (1 :1 ) and treated with LiOH (1 .05 equiv.) for half hour at room temperature. The reaction mixture was acidified to pH~2 with 1 N aqueous HCI. THF was removed under reduced pressure. The residue was extracted with EtOAc. After drying and condensation Cbz-Dab(Boc)-D-Thr-Dab(Boc)-OH was obtained in quantitative yield. Ions were found as: positive charge (M-Boc+H⁺: 554.2), negative charge (M-H^~: 653.2).

Step d. Synthesis of Dab(Boc)-D-Thr-Dab(Boc)-INT-11 : (2S)-2-[(N-{(2S)-2-

{[(benzyloxy)carbonyl]amino}-4-[(tert-butoxycarbonyl)amino]butanoyl}-D-threonyl)amino]-4-[(tert- butoxycarbonyl)amino]butanoic acid--tri-tert-butyl {[(2S,5R,8S,11 S,14S,17S,22S)-22- ({(8S,11 R,14S)-8-amino-14-{2-[(tert-butoxycarbonyl)amino]ethyl}-11-[(1S)-1-hydroxyethyl]-2,2- dimethyl-4,9,12,15-tetraoxo-3-oxa-5,10,13-triazapentadecan-15-yl}amino)-5-benzyl-17-[(1 R)-1- hydroxyethyl]-8-(2-methylpropyl)-3,6,9,12,15,18,23-heptaoxo-1 ,4,7,10,13,16,19- heptaazacyclotricosane-2,11 ,14-triyl]tri(ethane-2,1-diyl)}triscarbamate

INT-1 1 (0.709 g, 0.668 mmol) and Cbz-Dab(Boc)-D-Thr-Dab(Boc)-OH (0.502 g, 1 .577 mmol) were dissolved in 10 mL of DMF at room temperature followed by adding DIPEA (0.23 mL,1 .34 mmol). To this mixture was slowly added HATU (0.305 g, 0.801 mmol) in 1 .2 ml_ of DMF via syringe pump. After overnight of stirring at room temperature, the reaction mixture was condensed under reduced pressure. The residue was purified with reversed phase C18 with acetonitrile/water (0.1 %TFA) to give 1 .039 g of Cbz-protected Dab(Boc)-D-Thr-Dab(Boc)-INT-1 1 . Ion was found as ((M-2Boc+2H⁺)/2: 749.6). The Cbz protecting group was removed with 5% Pd/C in MeOH/EtOAc (1 :1 , 1 0 mL) under hydrogen balloon pressure at room temperature for 2 hours. Celite filtration gave Dab(Boc)-D-Thr-Dab(Boc)-INT-1 1 in quantitative yield. Ion was found as ((M+2H⁺)/2: 782.4).

Step e. Synthesis of Compound 23

The synthesis of Compound 23 was done in an analogous manner to Compound 21 by coupling

Dab(Boc)-D-Thr-Dab(Boc)-INT-1 1 with 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)diacetic acid (Example 23, Step d) followed by Boc deprotection with TFA. HPLC with ACN/water (0.1 % TFA) provided Compound 23 as a white powder after lyophilization. Ions were found as ((M+3H⁺)/3: 837.8).

Example 48. Synthesi

The title compound was prepared analogously to Compound 9 except the diacid of Example 31 , Step d was used as the linker, lon(s) found by LCMS: (M+2H)/2 = 1270.7, (M+3H)/3 = 847.5, (M+4H)/4 =635.9, (M+5H)/5 =508.9

Example 49. Prepara

The title compound was prepared analogously to Compound 24 but utilized INT-15 in place of INT-16 as the starting material, lon(s) found by LCMS: (M+3H)/3 = 891 .5, (M+4H)/4 =668.9, (M+5H)/5 =535.3

Example 50. Synthesis of INT-23

Step a. Synthesis of MeO-L-Dap-L-Thr-Cbz

MeO-L-Dap(p-Boc)-NH2 (HCI salt) (5.000g, 1 eq.), Z-NH-Thr-OH (5.049g, 1 .05eq.), EDCI (5.350g, 1 .5eq.), HOBt (3.733g, 1 .5eq.) and NaHC03 (3.095g, 2eq.) were weighed into a 100-mL round bottom flask. 24 mL of DCM/DMF (4:1 ) was added into the flask. The mixture was stirred at room temperature for less than 3 hrs. (TLC or LC/MS monitoring). After the completion, EtOAc (200 mL) was added to dilute the reaction mixture and washed with 1 N aq. HCI, saturated NaHC03 and brine. Dried with Na2S04 and condensed. The residue was purified with normal phase silica (2-7% MeOH/DCM) to give 8.24g pure desired product (>95%). Mass showed a strong detectable positive charge signal at t=4.1 07 min with 8 min (5-95%) (found M-Boc+H+: 368.21 ).

Step b. Synthesis of MeO-L-Dap-L-Thr-NH2

A solution of MeO-L-Dap-L-Thr-Cbz (1 .00g, 2.205 mmol) in methanol (1 OmL) was charged with 5%Pd/C (0.500 g), and hydrogen from a balloon. The reaction was monitored by LCMS. After 1 hr the mixture was filtered through Celite, concentrated and used immediately in the next step without purification.

Step c. Synthesis of MeO-L-Dap-L-Thr-L-Dab-Cbz

A solution of MeO-L-Dap-L-Thr-NH2(0.704g, 2.205mmol), (Ny-Boc)-L-Dab-Cbz (1 .295 g, 2.426 mmol), DIEA (1 .268 mL, 7.28 mmol), in DMF (8mL), was treated with a solution of HATU (0.922g,

2.426mmol) in DMF(4mL) dropwise via syringe pump over 1 hr. After 1 .5 hr the mixture was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield 1 .37 g, 95%. Step d. Synthesis of HO-L-Dap-L-Thr-L-Dab-Cbz

A solution of MeO-L-Dap-L-Thr-L-Dab-Cbz (1 .37g, 2.096mmol), in THF (5 mL) and water (5mL) was treated with LiOH (0.0552g, 2.310mmol) in one portion, at room temperature. LCMS after 15minutes show no starting material. The reaction was made slightly acidic with cone. HCI (aq), then extracted into ethyl acetate, and dried over sodium sulfate. The product was >90% pure by LCMS and was used without further purification, lon(s) found by LCMS: (M-H)- 638.2

Step e. Synthesis of ( Boc)- PM B H- L- Dap- L-Th - L- Dab-C bz

A solution of HO-L-Dap-L-Thr-L-Dab-Cbz (1 .190g, 1 .860mmol), Boc-PMB heptapeptide (Int 1 1 ,

1 .976 g, 1 .8603 mmol), DIEA (1 .07 mL, 6.14 mmol), in DMF (8 mL), was treated with a solution of HATU(0.778g, 2.046mmol) in DMF(2mL) dropwise via syringe pump over 1 hr. After 1 .5 hr the mixture was taken on to the next step without purification, lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 793.0, [(M- 2Boc)+2H]/2 = 743.0

Step f. Synthesis of (Boc)-PMBH-L-Dap-L-Thr-L-Dab-NH2

A solution of crude (Boc)-PMBH-L-Dap-L-Thr-L-Dab-Cbz (1 .86 mmol) in methanol (10mL) was charged with 5%Pd/C (1 .00 g), and hydrogen from a balloon. The reaction was monitored by LCMS. After 2 hr the mixture was filtered through Celite, concentrated and. purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 1 .43g. 49% . lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 725.9

Step g. Synthesis of (Boc)-PMBH-L-Dap-L-Thr-L-Dab-(L-amino-octanoyl-Cbz)

A solution of (Boc)-PMBH-L-Dap-L-Thr-L-Dab-NH2 (0.7g, 0.452mmol), Cbz-L-amino-octanoic acid(0.139g, 0.474mmol), and DIEA(0.248mL, 1 .423mmol) in DMF(5mL) was treated with a solution of HATU(0.180g, 0.474mmol) in DMF(1 mL) dropwise over 1 h. After 1 .5h the mixture was taken on to the next step without purification, lon(s) found by LCMS: (M+2H)/2 = 913.6, [(M-1 Boc)+2H]/2 = 863.6, [(M- 2Boc)+2H]/2 = 813.6

Step h. Synthesis of (Boc)-PMBH-L-Dap-L-Thr-L-Dab-(L-amino-octanoyl-NH2) (INT-23)

A solution of crude (Boc)-PMBH-L-Dap-L-Thr-L-Dab-(L-amino-octanoyl-Cbz) (0.452 mmol) was charged with 5%Pd/C (0.40 g), and hydrogen from a balloon. The reaction was monitored by LCMS. After 2 hr the mixture was filtered through Celite, concentrated and. purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.616g. 80% . lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 796.5, [(M-2Boc)+2H]/2 = 746.5, [(M-3Boc)+2H]/2 = 696.5 Example 51. Synthesis of INT-24

INT-24 was prepared analogously to INT-23 where MeO-L-Dap(p-Boc)-NH2 (HCI salt) was replaced with MeO-D-Dap(p-Boc)-NH2 (HCI salt) in the first step of that sequence, lon(s) found by LCMS: [(M-1 Boc)+2H]/2 = 796.5, [(M-2Boc)+2H]/2 = 746.5, [(M-3Boc)+2H]/2 = 696.5

Example 52. Synthesis of INT-25

INT-25 was prepared analogously to INT-23 where MeO-L-Dap(p-Boc)-NH2 (HCI salt) was replaced with MeO-L-Dab(p-Boc)-NH2 (HCI salt) in the first step of that sequence, lon(s) found by LCMS: [(M-2Boc)+2H]/2 = 753.0

Example 53. Synthesis of 3-{[(tert-butoxycarbonyl)(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]methyl}cyclopropane-trans-1 ,2-dicarboxylic acid (pair of diastereomers) (INT-26)

The aldehyde (dimethyl 3-formylcyclopropane-trans-1 ,2-dicarboxylate, racemic, prepared based on Reference Chem. Eur. J., 2013, 19, 6766-6773 , 0.785 g, 4.22 mmol) was dissolved in 20 mL of DCM. To this solution at room temperature, were added Rha-PEG1 -NH2 (INT-1 ) (1 .0596 g, 4.22 mmol, dissolved in 10 mL of 4:1 DCM/MeOH) and TFA (0.65 mL, 8.43 mmol). The mixture was stirred for 20 minutes followed by addition of NaBH(OAc)3 (1 .787 g, 8.43 mmol). The reaction mixture was stirred overnight to give the desired product (dimethyl 3-{[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]methyl}cyclopropane-trans-1 ,2-dicarboxylate, a pair of diastereomers) . Mass showed a strong detectable positive charge signal at tr=0.50 min with 8 min LC- MS run (found M+H⁺; 422.2). To this reaction mixture, were added DIPEA (2.20mL, 12.6 mmol) and Boc anhydride (2.03 mL, 8.40 mmol). After 3 hrs of stirring, the solvents were removed and the residue was purified with normal phase silica (2-7% MeOH/DCM) to give the desired product (dimethyl 3-{[(tert- butoxycarbonyl)(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)^

trans-1 ,2-dicarboxylate as a pair of diastereomers). Mass spectrum showed a strong detectable positive/negative charge signal at tr=3.60 min with 5 min polar run (2-95%) method (found Positive M- Boc+H⁺: 394.20; Negative M-H⁺; 492.00).

Example 54. Synthesis of INT-27

HATU (1 .56 g, 4.1 1 mmol) in DMF (1 .5 mL) was added, dropwise, to a solution of 2,2'- {[(benzyloxy)carbonyl]azanediyl}diacetic acid (0.5g, 1 .87 mmol), norleu-OMe hydrochloride salt (0.71 g, 3.93 mmol), and triethylamine (1 .13 g, 1 1 .23 mmol) in DMF (5 mL) over a period of 30 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the CBZ-protected di-ester as a clear oil, LC/MS [m+H]+ = 522.6. The di-ester intermediate was stirred in a 1 /1 /2 mixture (10 mL) of THF/methanol/water containing LiOH (0.1 8 g, 7.48 mmol) for 20 minutes. The mixture was applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford (2S,2'S)-2,2'-({[(benzyloxy)carbonyl]azanediyl}bis[(1 - oxoethane-2,1 -diyl)azanediyl])dihexanoic acid as a white solid. Yield: 43%, 2 steps. LC/MS [m-H]- =

492.3.

lnt-1 1 (500 mg, 0.47 mmol), HOBt (68 mg, 0.50 mmol), EDC (96 mg, 0.50 mmol), DIEA (130 mg, 1 mmol) and CBZ-(D)-Ser-OH (0.50 mmol) were stirred in DMF (3 mL) at room temperature for 2 hours. The mixture was applied directly to reversed phase HPLC (25-95% acetonitrile in Dl water containing 0.1 % TFA: 20 minute gradient). The pure fractions were pooled, concentrated and directly taken up in methanol (10 mL) and stirred in the presence of 5% Pd/C (75 mg) under 1 atmosphere of hydrogen gas for 1 hour. The mixture was filtered, concentrated and purified by reversed phase HPLC (10-95% acetonitrile in Dl water, no modifier: 20 minute gradient). The pure fractions were pooled and lyophilized to afford tri-tert-butyl {[(2S,5R,8S,1 1 S,14S,17S,22S)-5-benzyl-17-[(1 R)-1 -hydroxyethyl]-8-(2- methylpropyl)-3,6,9,12,15,18,23-heptaoxo-22-(D-serylamino)-1 ,4,7,10,13,16,19-heptaazacyclotricosane- 2,1 1 ,14-triyl]tri(ethane-2,1 -diyl)}triscarbamate (D-ser-(lnt-1 1 )) as a white solid. Yield: 83%. LC/MS [M- Boc+2H]/2 = 525.6.

Step b. Synthesis of L-thr-D-ser-(lnt-11)

L-threonyl-N-[(3S,6S,9S,12S,15R,18S.21 S)-15-benzyl-6,9,18-tris{2-[(tert- butoxycarbonyl)amino]ethyl}-3-[(1 R)-1 -hydroxyethyl]-12-(2-methylpropyl)-2,5,8,1 1 ,14,1 7,20-heptaoxo- 1 ,4,7,10,13,16,1 9-heptaazacyclotricosan-21 -yl]-D-serinamide (L-thr-D-ser-(lnM )) was prepared from tri- tert-butyl {[(2S,5R,8S,1 1 S,14S,17S,22S)-5-benzyl-17-[(1 R)-1 -hydroxyethyl]-8-(2-methylpropyl)- 3,6,9,12,15,18,23-heptaoxo-22-(D-serylamino)-1 ,4,7,10,13,16,19-heptaazacyclotricosane-2,1 1 ,14- triyl]tri(ethane-2,1 -diyl)}triscarbamate (D-ser-(lnt-1 1 )) and Z-Thr-OH using an analogous procedure to that described in Step a. Yield: 81 %. LC/MS [M-2 Boc+2H]/2 = 525.8.

Step c. Synthesis of Z-(Y-Boc)-Dab-L-thr-D-ser-(lnt-11)

INT-28 was prepared from the product of Step b and Z-(Y-Boc)-Dab-OH in an analogous manner as described in Step a. Yield: 78%. LC/MS [M+2H]/2 = 676.2 (loss of 1 Boc group).

Step d. Synthsis of INT-28

HATU (0.1 1 g, 0.30 mmol) in DMF (0.5 ml) was added, dropwise, to a stirring mixture of the product of Step c (0.44 g, 0.30 mmol), and INT-27 (0.70 g, 0.14 mmol) in DMF (1 .5 mL) over a period of 20 minutes. The mixture was applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 30% to 95% methanol and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the CBZ-protected intermediate, LC/MS [M+3H]/3 1020.0 (loss of 2 boc groups). The CBZ-intermediate was stirred in methanol (10 mL) in the presence of 5% Pd/C (100 mg) under 1 atmosphere of hydrogen for 2 hours. The mixture was filtered and concentrated then purified by to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 30% to 95% acetonitrile and water using no modifier. The pure fractions were pooled and lyophilized to afford the title compound as a white solid. Yield: 51 %, 2 steps. LC/MS [M+2H]/2 = 1562.6 (loss of 2 Boc groups). Example 56. Synthesis of INT-29

HATU (0.70 g, 1 .87 mmol) in DMF (1 ml_) was added, dropwise, to a solution of 1 -tert-butyl 4- methyl 4-aminopiperidine-1 ,4-dicarboxylate (0.48 g, 1 .87 mmol), CBZ-norleu-OH (0.45 g, 1 .70 mmol), and triethylamine (0.51 g g, 5.09 mmol) in DMF (2 ml_) over a period of 20 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the CBZ-protected intermediate as white solid. The intermediate was stirred under 1 atmosphere of hydrogen in the presence of 5% Pd/C (100 mg) for 2 hours. The mixture was filtered, concentrated, and applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford 1 -tert-butyl 4-methyl 4-(L-norleucylamino)piperidine-1 ,4-dicarboxylate as a white solid. Yield: 96%, 2 steps. LC/MS [m+H]+ = 372.3.

Example 57. Synthesis of INT-30

HATU (156 mg, 0.41 mmol) in DMF (1 mL) was added, dropwise, to a solution of INT-39 (144 mg, 0.41 mmol), INT-29 (265 mg, 0.32 mmol), and triethylamine (80 mg, 0.79 mmol) in DMF (5 mL) over a period of 30 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the di-ester as a clear oil, LC/MS [M+2H]/2 = 487.3 (loss of 2 boc groups). The di- ester intermediate was stirred in a 1 /1 /2 mixture (8 mL) of THF/methanol/water containing LiOH (30 mg, 1 .26 mmol) for 20 minutes. The mixture was applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 1 0% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford INT-30 as a white solid. Yield: 81 %, 2 steps. LC/MS [M+2H]/2 = 473.5 (loss of 2 boc groups).

Example 58. Synthesis of INT-31

HATU (156 mg, 0.41 mmol) in DMF (0.7 mL) was added, dropwise, to a solution of INT-30 (0.25 g, 0.22 mmol), H-Thr-(Y-Boc)Dab-OMe (0.16 g, 0.48 mmol), and triethylamine (1 10 mg, 1 .09 mmol) in DMF (1 .5 mL) over a period of 30 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 25% to 95% methanol and water using no modifier. The pure fractions were pooled and lyophilized to afford the di-ester as a clear oil, LC/MS [M+2H]/2 = 788.5 (loss of 2 boc groups). The di-ester intermediate was stirred in a 1 /1 /2 mixture (8 mL) of THF/methanol/water containing LiOH (31 mg, 1 .31 mmol) for 20 minutes. The mixture was applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 20% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford INT-31 as a white solid. Yield: 77%, 2 steps. LC/MS [M+2H]/2 = 774.6 (loss of 2 boc groups).

Example 59. Synthesis of INT-32

HATU (0.55 g, 1 .46 mmol) in DMF (1 mL) was added, dropwise, to a solution of INT-1 1 (1 .41 g, 1 .33 mmol), CBZ-Fluoro-Alanine-OH (0.35 g, 1 .46mmol), and triethylamine (0.40 g, 3.98 mmol) in DMF (2 mL) over a period of 20 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the CBZ-protected intermediate as white solid. The intermediate was stirred under 1 atmosphere of hydrogen in the presence of 5% Pd/C (100 mg) for 2 hours. The mixture was filtered, concentrated, and applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using no modifier. The pure fractions were pooled and lyophilized to afford the title compound as a white solid. Yield: 21 %, 2 steps. LC/MS [M+2H]/2 = 476.6 (loss of 2 Boc groups).

Example 60. Synthesis of INT-33

INT-33 was prepared from INT-32 and CBZ-Thr-OH by an analogous method as that for INT-32. Yield: 76%. LC/MS [M+2H]/2 = 527.0 (loss of 2 boc groups).

Example 61. Synthesis of INT-34

INT-34 was prepared from H-(Yboc)-Dab-OMe and CBZ-NorLeu-OH by a similar method as that for INT-32. Yield: 68%. LC/MS [m+H]+ = 246.4 (artifactual loss of 1 boc group).

Example 62. Synthesis of INT-35

HATU (6.3 g, 16.6 mmol) in DMF (2 mL) was added, dropwise, to a solution of 2,2'- {[(benzyloxy)carbonyl]azanediyl}diacetic acid (2.1 g, 7.9 mmol), INT-34 (5.7 g, 16.6 mmol), and triethylamine (4.0 g, 39.5 mmol) in DMF (4 mL) over a period of 20 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the CBZ-protected intermediate as white solid. The intermediate was stirred under 1 atmosphere of hydrogen in the presence of 5% Pd/C (500 mg) for 2 hours. The mixture was filtered, concentrated, and applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using no modifier. The pure fractions were pooled and lyophilized to afford the title compound as a white solid. Yield: 51 %, 2 steps. LC/MS [M+2H]/2 = 588.0 (loss of 2 boc groups).

Example 63. Synthesis of INT-36

INT-36 was prepared from INT-35 and INT-2 by a procedure analogous to the one for INT-30. Yield: 81 %, 2 steps. LC/MS [M-2H]/2 = 445.4 (loss of 2 Boc groups).

Example 64. Synthesis of INT-37a and INT-37b

HATU (3.1 g, 8.1 mmol) in DMF (5 mL) was added, dropwise, to a solution of racemic-transl - (ferf-butoxycarbonyl)pyrrolidine-3,4-dicarboxylic acid (1 g, 3.9 mmol), H-Norleu-OMe-hydrochloride (1 .5 g, 8.1 mmol), and triethylamine (4.0 g, 39.5 mmol) in DMF (10 mL) over a period of 20 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 1 0% to 95% acetonitrile and water using 0.1 % TFA modifier. The two diasteomers were separated and pooled and lyophilized separately into the more polar diasteomer (a) and the less polar diastereomer (b): LC/MS [m+H]+ = 414.2 (loss of 1 boc group) for both boc-protected intermediates. Each diastereomer was stirred in a 1 /1 mixture of DCM/TFA (8 mL) for 20 minutes then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 0% to 75% acetonitrile and water using no modifier. Yield: 88% (combined yield of both), 2 steps. LC/MS [M+H]+ = 414.2.

Note: The diastereomers have been arbitrarily assigned based on HPLC retention time. Example 65. Synthesis of INT-38a and INT-38b

INT-38a and INT-38b were prepared from INT-37a and INT-37b and 4-[(2-{2-[(6-deoxy-alpha-L mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid by a procedure analogous to the one for INT-30. Yield: 56%, 2 steps. LC/MS [m-H]- = 717.4.

Example 66. Synthesis of 2,2'-({4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoyl}azanediyl)diacetic acid (INT-39)

Step a. Synthesis of diethyl 2,2'-({4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoyl}azanediyl)diacetate

A mixture of 4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid (INT-2) (530 mg, 1 .51 mmol) and diethyl iminodiacetate (343 mg, 1 .82 mmol) was dissolved in anhydrous DMF (3 ml) and DIPEA (390 mg, 3 mmol). The solution was cooled in an ice-water bath and added with a solution of HATU (631 mg, 1 .66 mmol) in DMF (3 ml) via syringe pump at a rate of 1 .5 ml/hr. After the addition, the reaction was stirred for one more hour and then purified though RPLC (130 g, 5 to 15 % acetonitrile and water, using 0.1 % TFA as modifier). The collected fractions were concentrated by rotary evaporation and further dried under high vacuum to afford the title product as clear oil (680 mg, 86.2 %). Ion found by LCMS: [M]+ = 523.

Step b. Hydrolysis of diethyl 2,2'-({4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoyl}azanediyl)diacetate

The material from Step a was dissolved in a 1 :1 mixture of MeOH and THF (6 ml). The solution was cooled in an ice-water bath and added with a solution of LiOH (1 00 mg, 4 mmol) in water (3 ml). The resulting mixture was stirred at 0 °C to room temperature overnight. It was cooled back to an ice-water bath, and the pH was adjusted to ~ 7 by 4N HCI solution in dioxane. The organic solution was partially removed by rotary evaporation. The residue was purified through RPLC (150 g, 0 to 20 % MeOH and water, using 0.1 % TFA as modifier). Yield 564 mg, 93 % (INT-39). Ion found by LCMS: [M]+ = 467.

Example 67. Synthesis of INT-40

Step a. Synthesis of methyl N-{(2S)-2-{[(benzyloxy)carbonyl]amino}-4-[(tert- butoxycarbonyl)amino]butanoyl}-L-threoninate

A reaction flask was charged with a strong stirrer, L-threonine methyl ester HCI (2.4 g, 14 mmol) and anhydrous DMF (1 0 ml). The material was gently heated to dissolve. After cooled to room temperature, the solution was added with N^aZ-Nv-Boc-L-2,4-diaminobutyric acid dicyclohexylammonium (6.4 g, 12 mmol), DIPEA (3.64 g, 28 mmol), DMF (10 ml), and DCM (20 ml). The resulting suspension was added with a solution of HATU (4.8 g, 12.6 mmol) in DMF (12 ml) via syringe pump at a rate of 5 ml/hr. After the addition, the mixture was extracted with water (100 ml) and DCM (80 mL x 2). The combined organic layers were concentrated by rotary evaporation. The residue was purified through RPLC (150 g, 0 to 42 % acetonitrile and water, using 0.1 % TFA as modifier). Yield (5.25 g, 93.7 %). Ion found by LCMS: [M-Boc]+ = 368.

Step b. Synthesis of methyl N-{(2S)-2-amino-4-[(tert-butoxycarbonyl)amino]butanoyl}-L- threoninate

The step-a product (2.05 g, 4.28 mmol) was dissolved in MeOH (~ 20 ml). The solution was treated with Pd/C (1 g), and the resulting mixture was stirred under hydrogen for 2 hours. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation then further dried under high vacuum to a white solid. Yield (1 .41 g, 98.8 %). Ion found by LCMS: [M]+ = 334.

Step c. Synthesis of methyl (2S)-2-aminooctanoate

A suspension of (S)-2-aminooctanoic acid (650 mg, 4.08 mmol) in MeOH (8 ml) was cooled in an ice-water bath and added dropwise with thionyl chloride (0.5 ml). The mixture was warmed to room temperature and heated at 60 °C under nitrogen for 3 hours. It was then partially concentrated by rotary evaporation, and the residue was purified through RPLC (100 g, 0 to 25 % acetonitrile and water). The collected fractions were concentrated by rotary evaporation and further dried under high vacuum to afford the product as a clear oil (693 mg, 98 %). Ion found by LCMS: [M]+ = 174.

Step d. Synthesis of methyl (15S)-1-[(6-deoxy-alpha-L-mannopyranosyl)oxy]-15-hexyl-11-(2-{[(2S)-

1-methoxy-1-oxooctan-2-yl]amino}-2-oxoethyl)-7,10,13-trioxo-3-oxa-6,11 ,14-triazahexadecan-16- oate

A mixture of methyl (2S)-2-aminooctanoate (490 mg, 2.8 mmol) and 2,2'-({4-[(2-{2-[(6-deoxy- alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoyl}azanediyl)diacetic acid (600 mg, 1 .29 mmol) was dissolved in anhydrous DMF (2 ml) and DIPEA (702 mg, 5.4 mmol). The solution was cooled in an ice-water bath and drop-wise added with a solution of HATU (1 .06 g, 2.8 mmol) in DMF (3 ml) via syringe pump at a rate of 1 .5 ml/hr. After the addition, the reaction was directly purified through RPLC (100 g, 5 to 100 % acetonitrile and water). Yield 528.7 mg, 52.8 %. Ion found by LCMS: [M]+ = 777.

Step e. Synthesis of (15S)-11-(2-{[(1S)-1-carboxyheptyl]amino}-2-oxoethyl)-1-[(6-deoxy-alpha-L- mannopyranosyl)oxy]-15-hexyl-7,10,13-trioxo-3-oxa-6,11 ,14-triazahexadecan-16-oic acid

A solution of step-d product (528.7 mg, 0.681 mmol) in a 1 :1 mixture of MeoH:THF (6 ml) was cooled in an ice-water bath. It was added with a solution of LiOH (40 mg, 1 .67 mmol) in water (3 ml). The mixture was stirred for 3 hours, then neutralized by 4N HCI solution in dioxane. After partially concentrated, the reaction was directly purified through RPLC (100 g, 0 to 80 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 487.4 mg, 95.6 %. Ion found by LCMS: [M]+ = 749. Step f. Synthesis of INT-40-dimethyl ester

A mixture of the step-b product (481 mg, 1 .44mmol) and the step-e product (487.4 mg, 0.65 mmol) was dissolved in anhydrous DMF (2 ml) and DIPEA (338 mg, 2.6 mmol). The solution was cooled in an ice-water bath and added with a solution of HATU (532 mg, 1 .4 mmol) in DMF (2 ml) via syringe pump at a rate of 1 ml/hr. After the addition, the mixture was stirred for 30 minutes, then directly purified through RPLC (100 g, 1 0 to 100 % acetonitrile and water). Yield 735 mg, 81 .9 %. Ion found by LCMS: [(M-2Boc)/2]+ = 590.

Step g. Hydrolysis of INT-40-dimethyl ester

A solution of step-f product (735 mg, 0.533 mmol) in a 1 :1 mixture of MeOH:THF (10 ml) was cooled in an ice-water bath and treated with a solution of LiOH (40 mg, 1 mmol). The mixture was stirred for 3 hours, then neutralized by 4N HCI solution in dioxane. After partially concentrated, the mixture was purified through RPLC (150 g, 5 to 90 % MeOH and water, using 0.1 % TFA as modifier). Yield 712 mg, 98.8 %, white solid. Ion found by LCMS: [(M-2Boc)/2]+ = 576.

Example 68. Preparation of INT-41

Step a. Synthesis of 2,2'-(2-(benzyloxy)-2-oxoethylazanediyl)diacetic acid

Benzyl Bromo-acetate (4.6 g, 20 mmol) was added into the solution of 2,2'-azanediyldiacetic acid (2.7g, 20mmol) and DIPEA (5.6 g, 40 mmol) in 60 mL DMF, the resulted solution was stirred at room temperature for overnight. The reaction solution was concentrated and purified by flash chromatography to provide products. LC/MS, 282.1 [M+H]+.

Step b. Synthesis of benzyl 2-((2-oxo-2-(2-(2-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yloxy)ethoxy)ethylamino)ethyl)(2-oxo-2-(2-(2-((2R,3R,4R,5S)-3,4,5- trihydroxytetrahydro-2H-pyran-2-yloxy)ethoxy)ethylamino)ethyl)amino)acetate

To the solution of 2,2'-(2-(benzyloxy)-2-oxoethylazanediyl)diacetic acid (280mg, 1 mmol) in 5ml_

DMF were added (2R,3R,4R,5S)-2-(2-(2-aminoethoxy)ethoxy)tetrahydro-2H-pyran-3,4,5-triol (INT-1 ) (520mg, 2mmol), EDC (600mg, 3mmol), HOBT (420mg, 3mmol), Hunigs base (1 .4ml_, 10mmol) at room temperature. The solution was stirred for overnight. The resulted solution was removed DMF and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoacetic acid as the modifier. Yield of oil product, 520mg, 72% yield. LC/MS 734.3 [M+H]

Step c. Synthesis of 2-((2-oxo-2-(2-(2-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yloxy)ethoxy)ethylamino)ethyl)(2-oxo-2-(2-(2-((2R,3R,4R,5S)-3,4,5-trihydroxytetrahydro- 2H-pyran-2-yloxy)ethoxy)ethylamino)ethyl)amino)acetic acid

Benzyl 2-((2-oxo-2-(2-(2-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yloxy)ethoxy)ethylamino)ethyl)(2-oxo-2-(2-(2-((2R,3R,4R,5S)-3,4,5-trihydroxytetrahydro-2H-pyran-2- yloxy)ethoxy)ethylamino)ethyl)amino)acetate (500mg, 0.7mmol) was dissolved into 5mL MeOH and 3 mL ethyl acetate, then 200mg of 5% Palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for overnight. The palladium charcoal was removed by filtration after completed the reaction by LCMS. The filtrate was concentrated and used next step without any purification. LC/MS, 644.3 [M+H]+ . Step d. Synthesis of (S)-benzyl 17-(2-(bis(2-oxo-2-(2-(2-((2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yloxy)ethoxy)ethylamino)ethyl)amino)acetamido)-7,11-dioxo-9- oxo-2-(2-(2-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yloxy)ethoxy)ethylamino)ethyl)-1-((2R,3R,4R,5R,6S)-3,4 trihydroxy-6-methyltetrahydro-2H-pyran- 2-yloxy)-3-oxa-6,9,12-triazaoctadecan-18-oate

To the solution of 2-((2-oxo-2-(2-(2-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yloxy)ethoxy)ethylamino)ethyl)(2-oxo-2-(2-(2-((2R,3R,4R,5S)-3,4,5-trihydroxytetrahydro-2H- pyran-2-yloxy)ethoxy)ethylamino)ethyl)amino)acetic acid (400mg, 0.6mmol) in 5ml_ DMF were added (S)-benzyl 2,6-diaminohexanoate (70mg, 0.3mmol), EDC (200mg, 1 mmol), HOBT (150mg, 1 mmol), Hunigs base (0.28mL, 2mmol) at room temperature. The solution was stirred for overnight. The resulted solution was removed DMF and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 1 00% acetonitrile and water, using 0.1 % trifluoacetic acid as the modifier. Yield of oil product, 360mg, 78% yield. LC/MS 758.4 [(M+2H)/2].

Step e. Synthesis of (S)-17-(2-(bis(2-oxo-2-(2-(2-((2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yloxy)ethoxy)ethylamino)ethyl)amino)acetamido)-7,11-dioxo-9-(2- oxo-2-(2-(2-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2- yloxy)ethoxy)ethylamino)ethyl)-1-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran- 2-yloxy)-3-oxa-6,9,12-triazaoctadecan-18-oic acid (INT-41)

(S)-benzyl 17-(2-(bis(2-oxo-2-(2-(2-((2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6-methyltetrahydro-2H- pyran-2-yloxy)ethoxy)ethylamino)ethyl)amino)acetamido)-7,1 1 -dioxo-9-(2-oxo-2-(2-(2-((2R,3R,4R,5R,6S)- 3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yloxy)ethoxy) ethylamino)ethyl)-1 -((2R,3R,4R,5R,6S)- 3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yloxy)-3-oxa-6,9,12-triazaoctadecan-18-oate (300 mg, 0.2 mmol) was dissolved into 2ml_ MeOH and 2 ml_ ethyl acetate, then 100mg of 5% Palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for overnight. The palladium charcoal was removed by filtration after completed the reaction by LCMS. The filtrate was concentrated and used next step without any purification. LC/MS, 713.7

[(M+2H)/2] . Exam le 69. Synthesis of Compound 26

Step a. Synthesis of methyl (2S)-2-[(2-{[(benzyloxy)carbonyl]amino}ethyl)amino]-4-[(fert- butoxycarbonyl)amino]butanoate

To a solution of (S)-methyl-2-amino-4-((tert-butoxycarbonyl)amino)butanoate HCI (537.5 mg, 2 mmol) in DMF/water (2/0.5 ml) was added benzyl 2-bromoethylcarbamate (567.8 mg, 2.2 mmol) and sodium bicarbonate (252 mg, 3 mmol). After the resulting mixture was stirred for 3 hours, it was added with an additional amount of benzyl 2-bromoethylcarbamate (258 mg, 1 mmol). The reaction was continued overnight and purified through RPLC (1 50 g, 5 to 100 % acetonitrile and water). The collected fractions were concentrated by rotary evaporation to afford the title compound as a clear oil (750 mg, 91 .6%). Ion found by LCMS: [M]+ = 410.

Step b. Synthesis of (8S,11 £,15S)-9,14-bis(2-{[(benzyloxy)carbonyl]amino}ethyl)-15-{2-[(fert- butoxycarbonyl)amino]ethyl}-8-carboxy-2,2-dimethyl-4,10,13-trioxo-3-oxa-5,9,14-triazahexadec-11- en-16-oic acid

A flame-dried reaction flask was filled with nitrogen and charged with step-a product (328 mg, 0.8 mmol), chloroform (2 ml) and DIPEA (41 6 mg, 3.2 mmol). After the solution was cooled in an ice-water bath, it was syringe-pumped added with a solution of fumaryl chloride (61 mg, 0.4 mmol) in chloroform (0.6 ml) at a rate of 0.2 ml/hr. An additional amount of DIPEA (208 mg, 1 .6 mmol) was added for every hour. After the completion of the fumaryl chloride addition, the reaction was stirred for one more hour and then concentrated by rotary evaporation. The residue was purified through RPLC (50 g, 10 to 70 % acetonitrile and water, using 0.1 % TFA as modifier). The collected fractions were concentrated by rotary evaporation to a purple solid (LCMS: [M]+ = 899). The material was re-dissolved in 1 :1 MeOH:THF (3 ml). After cooled in an ice-water bath, the solution was added with LiOH solution (20 mg, 0.8 mmol) in water (1 .5 ml). The mixture was stirred for 3 hours and then acidified by formic acid. It was purified through RPLC (50 g, 20 to 78 % MeOH and water). The collected fractions were concentrated by rotary evaporation and further dried under high vacuum to afford the title product as a white solid (156 mg, 22.4 %). Ion found by LCMS: [M-Boc]+1 = 771 .

Step c. Synthesis of A ^, ,Af-bis(2-aminoethyl)butanediamide-INT-16

To a mixture of step-b product (56 mg, 0.0643 mmol), INT-16 (192.9 mg, 0.141 mmol) and DIPEA

(78 mg, 0.6 mmol) in anhydrous DMF (1 ml) was added with a solution of HATU (26.9 mg, 0.07 mmol) in DMF (1 ml) via syringe pump at 0.5 ml/hr rate. After the addition, the mixture was stirred for one more hour and then added with Pd/C and EtOAc. The resulting mixture was stirred under hydrogen for 4 hours. Pd/C was filtered off, and the filtrate was concentrate. The residue was purified through RPLC (50 g, 20 to 100 % MeOH and water, using 0.1 % formic acid as modifier). The collected fractions were

concentrated by rotary evaporation to a white solid (34.5 mg, 16.3%). Ions found by LCMS: [(M-Boc)/3]+1 = 1 065, [(M-3Boc)+3H]/3 = 999.

Step d. Synthesis of deca-Boc-Compound 26 precursor

To a solution of a mixture of 4-[(2-{2-[(6-deoxy-a -L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoic acid (INT-2) (10 mg, 0.0285 mmol) and HOBT hydrate (5.7 mg, 0.037 mmol) in anhydrous DMF (0.5 ml) was added EDC-HCI (6.1 mg, 0.032 mmol). After stirred for 10 minutes, it was added with a solution of step-c product (34.5 mg, 0.0105 mmol) and DIPEA (50 mg, 0.385 mmol) in DMF (1 ml). The reaction was stirred for 2 hours, then directly purified through RPLC (50g, 20 to 100 % MeOH and water). The collected fractions was concentrated by rotary evaporation to a white solid (24.9 mg, LCMS [(M- Boc)/3]+1 = 1287, [(M-2Boc)+3H]/3 = 1254).

Step e. Removal of the Boc group

The step-d product was re-dissolved in TFA/DCM (1 :1 , - 1 ml). After the solution was stirred for 15 minutes, it was purified through HPLC (5 to 22 % acetonitrile and water, using 0.1 % TFA as modifier). Yield (5.8 mg, 13.5 %). Ions found by LCMS: (M+3H)/3 = 987, (M+4H)/4 = 740, (M+5H)/5 = 592. Example 70. Synthesis of Compound 27

HATU (40 mg, 0.106 mmol) in DMF (0.5 ml) was added, dropwise, to a stirring mixture of INT-28 (0.23 g, 0.071 mmol), and 4-[(2-{2-[(6-deoxy-alpha-L mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoic acid (INT-2) (38 mg, 0.107 mmol) in DMF (1 .5 mL) over a period of 20 minutes. The mixture was applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 30% to 95% methanol and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the boc-protected intermediate; LC/MS [M+3H]/3 1086.6 (loss of 2 boc groups). The boc-intermediate was stirred in a 1 /1 mixture of DCM/TFA (5 mL) containing thioanisole (71 mg, 0.071 mmol) for 20 minutes. The mixture was concentrated on the rotovap then applied to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 0% to 55% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford the title compound as a white solid. Yield: 41 %, 2 steps. LC/MS [M+3H]/3 = 919.8.

Step a. Coupling of INT-20

To the solution of 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}azanediyl)diacetic acid (1 0 mg, 0.023 mmol), triethyl amine (0.07 mL, 0.5 mmol) and INT-20 (85 mg, 0.05 mmol) in 5ml_ DMF was added HATU (40 mg, 0.1 mmol) in 1 mL DMF by syringe pump over 1 hour. Then the resulted solution was concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of product, 60mg, 70% yield.

Step b. Removal of the Boc groups

The material from Step a was treated in 2 mL DCM and 2mL TFA at room temperature, the solution was stirred 10 min, then concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 35 mg, 60% yield (Compound 28). lon(s) found by LCMS:

(M+3H)/3 = 932.9, (M+4H)/4 =699.9, (M+5H)/5=560.1 .

Step a. Coupling to INT-20

To the solution of (S)-3-(carboxymethyl)-7-(octylcarbamoyl)-5,9-dioxo-1 5-(((2R,3R,4R,5R,6S)- 3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-13-oxa-3,6,10-triazapentadecan-1 -oic acid (15 mg, 0.023 mmol), triethyl amine (0.07mL, 0.5mmol) and INT-20 (85mg, 0.05mml) in 5mL DMF was added HATU (19 mg, 0.5mmol) in 1 mL DMF by syringe pump over 1 hour. Then the resulted solution was concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoacetic acid as the modifier. Yield of product, 45mg, 51 % yield.

Step b. Removal of the Boc groups

The per-Boc product from Step a was treated in 2 mL DCM and 2mL TFA at room temperature, the solution was stirred 10 min, then concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 15mg, 48% yield (Compound 29). lon(s) found by LCMS:

(M+3H)/3 = 1008.3, (M+4H)/4 =756.5, (M+5H)/5=605.4, (M+6H)/6=504.6.

HATU (78 mg, 0.20 mmol) in DMF (0.5 mL) was added, dropwise, to a solution of INT-31 (0.17 g, 0.097 mmol), trisBoc-PMBH (INT-1 1 ) (227 mg, 0.21 mmol), and triethylamine (59 mg, 0.58 mmol) in DMF (1 .5 mL) over a period of 30 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 25% to 95% methanol and water using no modifier. The boc-protected intermediate was stirred in a 1 /1 mixture of DCM/TFA (4 mL) containing thioanisole (121 mg, 0.97 mmol) for 20 minutes. The solvent was removed by rotovap and the residue was dried under high vacuum and then purified by reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 0% to 45% acetonitrile and water using no modifier. The pure fractions were pooled and lyophilized to afford the title compound as a white solid. Yield: 59%, 2 steps. LC/MS [M+3H]/3 = 945.9.

Step a. Synthesis of deca-Boc Compound 31 benzyl ester

Deca-Boc Compound 31 benzyl ester was prepared analogously to Example 37 (alternate procedure), where racemic (1 R,2R)-3-[(benzyloxy)carbonyl]cyclopropane-1 ,2-dicarboxylic acid, was substituted for 2,2'-{[(benzyloxy)carbonyl]azanediyl}diacetic acid in the first step of the sequence.

Step b. Synthesis of deca-Boc Compound 31 carboxylic acid

Crude deca-Boc benzyl ester Intermediate (0.373g, 0.103 mmol) from Step a (DMF solution) was charged with 5%Pd/C (0.200 g), and hydrogen from a balloon. The reaction was monitored by LCMS. After 12 hr the mixture was filtered through Celite, concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.295 g, 81 % (two steps), lon(s) found by LCMS: [(M- 3Boc)+3H]/3 = 1083.0, [(M-4Boc)+3H]/3 = 1049.7, [(M-5Boc)+3H]/3 = 1016.3

Step c. Synthesis of deca-Boc Compound 31

A solution of deca-Boc Compound 31 carboxylic acid (0.295g, 0.0831 mmol), L-Rhamnose-PEG1 - NH2 (INT-1 ) (0.01 15g, 0.0457mmol), DIEA(0.0239DL, 0.137mmol), in DMF(3mL), was treated with a solution of HATU(0.0174g, 0.0457mmol) in DMF(0.5mL) dropwise via syringe pump over 1 hr. After 1 .5 hr the mixture was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.057 g, 36%. lon(s) found by LCMS: (M+3H)/3 = 1260.7, [(M-1 Boc)+3H]/3 = 1227.4, [(M-2Boc)+3H]/3 = 1 1 94.0 Step d. Synthesis of Compound 31

A solution of deca-Boc-Compound 31 (0.234 g, 0.0659 mmol), dissolved in DCM (1 ml_), was treated with TFA (1 ml_), while stirring at room temperature. After 5 minutes, the product was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 0.027 g, 55% yield (mixture of diastereomers). lon(s) found by LCMS: (M+3H)/3 = 927.2, (M+4H)/4 = 695.7, (M+5H)/5 = 556.7, (M+6H)/6 = 464.1 , (M+7H)/7 = 398.0

Compound 32 was prepared analogously to example 37 (alternate procedure) where intermediate INT-20 was replaced with intermediate INT-23 in the first step of that sequence, lon(s) found by LCMS: (M+3H)/3 = 938.1 , (M+4H)/4 = 703.8, (M+5H)/5 = 563.3

Example 76. Synthesis of Compound 33

Compound 33 was prepared analogously to Example 75 where INT-23 was replaced with INT-24 in the first step of that sequence, lon(s) found by LCMS: (M+4H)/4 = 703.8, (M+5H)/5 = 563.3, (M+6H)/6 = 469.6, (M+7H)/7 = 402.6

Step a. Synthesis of ethyl 32-(2-ethoxy-2-oxoethyl)-3,31 -dioxo-1 -phenyl-2,7,10,13,16,19,22,25,28- nonaoxa-4,32-diazatetratriacontan-34-oate

To the solution of 3-oxo-1 -phenyl-2,7,10,13,16,19,22,25,28-nonaoxa-4-azahentriacontan-31 -oic acid (1 g, 2.5 mmol) in DMF (20 mL) were added diethyl 2,2'-azanediyldiacetate (0.71 g, 3.7 mmol), EDC (600mg, 3mmol), HOBT (0.45g, 3.0mmol), Hunigs base (0.7 mL, 5mmol) at room temperature. The solution was stirred overnight. The resultant solution was removed DMF and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of oil product, 1 .2 g, 65% yield. LC/MS 747.4 [M+H]

Step b. Synthesis of 32-(carboxymethyl)-3,31-dioxo-1 -phenyl-2,7,10,13,16,19,22,25,28-nonaoxa- 4,32-diazatetratriacontan-34-oic acid

Lithium hydroxide (50 mg, 2 mmol) in 2 mL H2O was added to the solution of ethyl 32-(2-ethoxy- 2-oxoethyl)-3,31 -dioxo-1 -phenyl-2,7,10,13,16,19,22,25,28-nonaoxa-4,32-diazatetratriacontan-34-oate (800mg, 1 mmol) in mix solvent of 2 mL H2O, 2mL MeOH and 2mL THF. The resulting solution was stirred for 1 hour at room temperature. The resulted solution was concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoro-acetic acid as the modifier. LC/MS 691 .3 [M+H]⁺

Step c. Synthesis of Bis (penta-Boc-polymyxin B decapeptide)-32-(carboxymethyl)-3,31 -dioxo-1 - phenyl-2,7,10,13,16,19,22,25,28-nonaoxa-4,32-diazatetratriacontan-34-amide

To a solution of 32-(carboxymethyl)-3,31 -dioxo-1 -phenyl-2,7,10,13,16,19,22,25,28-nonaoxa-4,32- diazatetratriacontan-34-oic acid (70 mg, 0.1 mmol), triethyl amine (0.14 mL, 1 mmol) and INT-18 (300 mg, 0.2 mmol) in 5 mL DMF was added HATU (80 mg, 0.2 mmol) in 1 mL DMF by syringe pump over 1 hour. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of product, 230 mg, 70% yield.

Step d. Synthesis of deca-Boc-lntermediate

The Cbz-protected product from Step c (120 mg, 0.03 mmol) was dissolved into a mixture of 2 mLs of MeOH and 2 mLs of ethyl acetate, then 100 mg of 5% palladium on charcoal was added to the above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere overnight. The palladium charcoal was removed by filtration after completion of the reaction as indiciated by HPLC. The filtrate was concentrated and used next step without any purification.

Step e. Coupling to INT-41

To the solution of (S)-17-(2-(bis(2-oxo-2-(2-(2-((2S,3S,4S,5S,6R)-3,4,5-trihydroxy-6- methyltetrahydro-2H-pyran-2-yloxy)ethoxy)ethylamino)ethyl)amino)acetamido)-7,1 1 -dioxo-9-(2-oxo-2-(2- (2-((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yloxy) ethoxy)ethylamino)ethyl)-1 - ((2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yloxy)-3-oxa-6,9,12-triazaoctadecan- 18-oic acid (27mg, 0.02 mmol), triethyl amine (0.07mL, 0.5mmol) and INT-41 (70 mg, 0.02 mmol) in 5 mL DMF was added HATU (19 mg, 0.05 mmol) in 1 mL DMF by syringe pump over 1 hour. Then the resulted solution was concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco

CombiFlash liquid chromatograph eluted with 5% to 1 00% acetonitrile and water, using 0.1 % trifluoacetic acid as the modifier. Yield of product, 60mg, 55% yield.

Step f. Removal of the Boc groups

The product from Step e was treated in 2 mL DCM and 2 mL TFA at room temperature, the solution was stirred 10 min, then concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 16 mg, 38% yield (Compound 34). lon(s) found by LCMS:

(M+4H)/4 = 1014.1 , (M+5H)/5 =81 1 .4, (M+6H)/6=676.4, (M+6H)/6=579.9.

Example 78. Synthesis of Compound 35

Step a. Synthesis of Met-INT-18

A reaction flask was filled with nitrogen and charged with a mixture of Fmoc-L-Met-OH (44.6 mg,

0.12 mmol) and HOBT hydrate (26 mg, 0.17 mmol) and anhydrous DMF ((1 ml). EDC-HCI (25.2 mg, 0.132 mmol) was added, and the resulting mixture was stirred for 15 minutes, then added with INT-18 (168 mg, 0.1 mmol) and DIPEA (91 .5 mg, 0.7 mmol). After stirred for 1 hour, the reaction mixture was quenched with water (0.1 ml) and stirred for 10 minutes. Piperidine (3 drops) was then added, and the resulting mixture was stirred for 2 hours. It was then directly purified through RPLC (50 g, 20 to 82 % MeOH and water). Yield 139.6 mg, 82.4 %. Ions found by LCMS: [(M-Boc)/2]+1 = 798.

Step b. Synthesis of decaBoc-Compound 35 precursor

A reaction flask was filled with nitrogen and charged with a mixture of 2,2'-({4-[(2-{2-[(6-deoxy- alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoyl}azanediyl)diacetic acid (1 1 mg, 0.0236 mmol), step-a product (84.8 mg, 0.05 mmol), anhydrous DMF (0.5 ml), and DIPEA (39 mg, 0.3 mmol). The mixture was cooled in an ice-water bath and added with a solution of HATU (19 mg, 0.05 mmol) in DMF (0.5 ml) via syringe pump at a rate of 0.5 ml/hr. After the addition, the reaction was stirred for one more hour. It was then purified through RPLC (50 g, 20 to 100 % MeOH and water). Yield 53.7 mg, 40.4 %. Ions found by LCMS: [(M-Boc)/3]+1 = 1240, [(M-2Boc)/3]+= 1207, [(M-3Boc)/3]+= 1 173.

Step c. Removal of the Boc groups

The step-b product was dissolved in TFA (-0.5 ml). The solution was stirred at room temperature for 15 minutes, then directly purified through HPLC (5 to 20 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 20.4 mg, %. Ions found by LCMS: [M/3]+ = 940, [M/4]+ = 705, [M/5]+ = 564, [M/6]+ = 470.

Example 79. Synthesis of Compound 36

The title compound was prepared analogously to Compound 35. Ions found by LCMS: [M/3]+ 950, [M/4]+ = 713, [M/5]+ = 570, [M/6]+ = 475.

Example 80. Synthesis of Compound 37

The title compound was prepared analogously to Compound 35. Ions found by LCMS: [M/3]+ 961 , [M/4]+ = 721 , [M/5]+ = 577, [M/6]+ = 481 .

Example 81. Synthesis of Compound 38

The title compound was prepared analogously to Compound 35. Ions found by LCMS: [M/4]+ 698, [M/5]+ = 558, [M/6]+ = 466, (M+7H)/7= 399.

Example 82. Synthesis of Compound 39

The title compound was prepared analogously to Compound 35 Ions found by LCMS: [M/3]+ 940, [M/4]+ = 705, [M/5]+ = 564, [M/6]+ = 470, [M/7]+ = 403.

Example 83. Synthesis of Compound 40

Compound 40 was prepared analogously as the compound of Example 37 (alternate procedure) where intermediate INT-20 was replaced with INT-25 in the first step of that sequence, lon(s) found by LCMS: (M+3H)/3 = 946.9, (M+4H)/4 = 710.4, (M+5H)/5 = 568.5, (M+6H)/6 = 473.9, (M+7H)/7 = 406.4 Example 84. Synthesis of Compound 41

Step a. Synthesis of Ai-{[(9W-fluoren-9-yl)methoxy]carbonyl}-S-propyl-L-homocysteine

A solution of NaOH (600 mg, 15 mmol) in 1 :1 mixture of water/EtOH (10 ml) was bubbled with nitrogen and heated at 40 °C. It was added with L-homocysteine (675.5 mg, 5 mmol), followed by propyl p-toluenesulfonate (1 .09 g, 5.05 mmol). The mixture was stirred under nitrogen at 40 °C for 1 .5 hours, then to room temperature overnight. EtOH was partially removed by rotary evaporation, and the residue was extracted with DCM (~ 30 ml). THF (10 ml) was added to the aqueous solution, followed by Fmoc- Osu (1 .7 g, 5 mmol). After the mixture was stirred for 1 hour, it was added with an additional amount of Fmoc-Osu (337 mg, 1 mmol). The reaction was continued for one more hour. THF was partially removed by rotary evaporation. The white suspension residue was directly purified through RPLC (150 g, 5 to 58 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 1 .05 g, 50.3 %. Ion found by LCMS: [M+1 ]+ = 400.

Step b. Synthesis of (propyl)-homocys-INT-18

The title compound was prepared analogously to Met- INT- 18. Ion found by LCMS: [(M-Boc)/2]+

= 812. Step c. Synthesis of Compound 41

The title compound was prepared analogously to Compound 35 Ions found by LCMS: [M/3]+ 959, [M/4]+ = 719, [M/5]+ = 575, [M/6]+ = 480.

Example 85. Synthesis of Compound 42

Step a. Synthesis of (cyclopropyl)-ala-INT-18

A mixture of Fmoc-L-cyclopropylalanine (42.2 mg, 0.12 mmol) and HOBT hydrate (25.7 mg, 0.168 mmol) was dissolved in an anhydrous DMF (0.5 ml). EDC-HCI (25.2 mg, 0.132 mmol) was added, and the resulting mixture was stirred for 15 minutes. It was added with INT-18 (168 mg, 0.1 mmol), followed by DIPEA (130 mg, 1 mmol). After stirred for 1 hour, the reaction mixture was quenched with water (0.1 ml) and stirred for 10 more minutes. Piperidine (3 drops) was then added, and the resulting mixture was stirred for 2 hours. It was then directly purified through RPLC (50 g, 20 to 100 % MeOH and water). Yield 145.7 mg, 87 %. Ion found by LCMS: [(M-Boc)/2]+1 = 788. Step b. Synthesis of decaBoc-Compound 42 precursor

A mixture of 2,2'-({4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoyl}azanediyl)diacetic acid (15.7 mg, 0.0336 mmol) and the step-b product (1 15 mg, 0.0687 mmol) was dissolved in anhydrous DMF (0.5 ml) and DIPEA (39 mg, 0.3 mmol). The solution was cooled in an ice-water bath and added with a solution of HATU (28 mg, 0.074 mmol) in DMF (0.5 ml) via syringe pump at a rate of 0.5 ml/hr. After the addition, the reaction was stirred for one more hour and directly purified through RPLC (50 g, 20 to 100 % MeOH and water). Yield 73.2 mg, 57.6 %. Ions found by LCMS: [(M-Boc)/3]+ = 1227, [(M-2Boc)+3H]/3 = 1 193, [(M-3Boc)/3]+= 1 160, [(M-5Boc)/3]+= 1093.

Step c. Removal of the Boc groups

DecaBoc-Compound 42 precursor was dissolved in TFA (-0.5 ml). The solution was stirred at room temperature for 15 minutes, then directly purified through HPLC (5 to 20 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 20.4 mg, 20.4 %. Ions found by LCMS: [M/3]+ = 926, [M/4]+ = 695, [M/5]+ = 556, [M/6]+ = 463.

Example 86. Synthesis of Compound 43

Step a. Synthesis of L-Thre-IN -18

To a mixture of Z-thr-OH (30.4 mg, 0.12 mmol) and HOBT hydrate (25.7 mg, 0.168 mmol) was dissolved in anhydrous DMF (0.5 ml) was added EDC-HCI (25.2 mg, 0.132 mmol). The resulting mixture was stirred for 15 minutes, then added with INT-18 (168 mg, 0.1 mmol), followed by DIPEA (130 mg, 1 mmol). After stirring for 1 hour, the reaction mixture was purified through RPLC (50 g, 20 to 1 00 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (LCMS: [(M- 2B0c)/2]+ = 800). The material was re-dissolved in MeOH (~ 20 ml) and added with Pd/C. The resulting mixture was stirred under hydrogen for 2 hours. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried under high vacuum. Yield 120 mg, 72 %. Ion found by LCMS: [(M- Boc)/2]+1 = 783.

Step b. Synthesis of decaBoc-Compound 43 precursor

The title compound was prepared analogously to decaBoc-Compound 42 precursor. Ions found by LCMS: [(M-Boc)/3]+ = 1220, [(M-2Boc)+3H]/3 = 1 1 87, [(M-3Boc)/3]+= 1 153.

Step c. Removal of the Boc group

The reaction was performed analogously to the removal of the Boc group of deca-Boc Compound 42 precursor. Ions found by LCMS: [M/5]+ = 552, [M/6]+ = 460, [M/7]+ = 395. Example 87.

Compound 44 was prepared from INT-33 and INT-36 as described in the procedure for the synthesis of Compound 30. Yield: 15%, 2 steps. LC/MS [M+3H]/3 = 921 .0.

Example 88. Synthesis of Compound 45

Step a. Coupling of INT-18

To the solution of 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}azanediyl)diacetic acid (1 0mg, 0.023 mmol), triethyl amine (0.07ml_, 0.5mmol) and INT-18 (74mg, 0.047mml) in 5ml_ DMF was added HATU(40mg, O.o2mmol) in 1 mL DMF by syringe pump over 1 hour. Then the resulted solution was concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoacetic acid as the modifier. Yield of product, 30mg, 37% yield.

Step b. Removal of Boc groups

The product from Step a was treated in 2 ml_ DCM and 2ml_ TFA at room temperature, the solution was stirred 10 min, then concentrated and purified by reverse phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using formic acid as the modifier. Yield 15mg, 70% yield, lon(s) found by LCMS: (M+2H)/2 = 1298.8, (M+3H)/3 = 866.2, (M+4H)/4 =649.9, (M+5H)/5=519.1 .

Example 89. Synthesis of Compound 46

Step a. Synthesis of (2¾-2-({[(9W-fluoren-9-yl)methoxy]carbonyl}amino)-5-methylhex-4-enoic acid

c

(S)-2-amino-5-methylhex-4-enoic acid (715.9 mg, 5 mmol) was dissolved in water (3 ml) by gently heating with a heat-gun. After cooled to room temperature, the solution was added with THF (6 ml) and a solution of NaOH (370 mg, 9.25 mmol) in water (3 ml). Fmoc-Osu (1 .7 g, 5.05 mmol) was added, and the resulting mixture was stirred for 1 .5 hours. It was then directly purified through RPLC (150 g, 5 to 60 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 598.6 mg, 32.7%. Ion found by LCMS: [M+1 ]+ = 366. Step b. Synthesis of 5-methyl

The title compound was prepared analogously to (cyclopropyl)-ala-INT-l 8. Ion found by LCMS: [(M-Boc)/2]+ = 795.

Step c. Synthesis of Deca-Boc Compound 46 precursor

A mixture of 2,2'-({4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoyl}azanediyl)diacetic acid (18 mg, 0.0385 mmol), 5-methylhex-INT-18 (143.5 mg, 0.085 mmol), DIPEA (42 mg, 0.4 mmol) and anhydrous DMF (1 ml). It was cooled in an ice-water bath and added with a solution of HATU (32.2 mg, 0.0847 mmol) in DMF (0.5 ml) via syringe pump at a rate of 0.5 ml/hr. After the addition, the reaction was stirred for one more hour. It was then purified through RPLC (50 g, 20 to 100 % MeOH and water). Yield 1 03.4 mg, 70.5 %. Ions found by LCMS: [(M-Boc)/3]+1 = 1236, [(M- 2Boc)/3]+= 1203, [(M-3Boc)/3]+= 1 170.

Step d. Removal of the Boc groups

Deca-Boc Compound 46 precursor product (50 mg, 0.013 mmol)) was dissolved in a 1 :1 mixture of TFA:DCM (-0.5 ml). The solution was stirred at room temperature for 15 minutes, then directly purified through HPLC (5 to 20 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 10.4 mg,

20.3%. Ions found by LCMS: [M/3]+ = 935 , [M/4]+ = 702, [M/5]+ = 562, [M/6]+ = 468. Example 90. Synthesis of Compound 47

Deca-Boc Compound 46 precursor (53 mg, 0.138 mmol) was dissolved in MeOH (~ 5 ml). The solution was added with 1 0% Pd/C, and the mixture was stirred under hydrogen overnight. Pd/C was filtered off, and the filtrated was concentrated by rotary evaporation to a white solid (44 mg, 0.01 15 mmol, 83 %). The material was dissolved to a a 1 :1 mixture of TFA:DCM which was precooled in an ice-water bath. The solution was stirred at 0°C to room temperature for 1 hour. It was then directly purified through HPLC. Yield 23.2 mg, 51 .3%. Ions found by LCMS: [M/3]+ = 937 , [M/4]+ = 703, [M/5]+ = 562, [M/6]+ = 469.

Example 91. Synt

Compound 48 was prepared analogously to example 37 (alternate procedure) where 2,2'- {[(benzyloxy)carbonyl]azanediyl}diacetic acid was replaced with N-[(benzyloxy)carbonyl]-D-aspartic acid in the first step of that sequence, lon(s) found by LCMS: (M+3H)/3 = 852.8, (M+4H)/4 = 639.9, (M+5H)/5 = 512.1 , (M+6H)/6 = 427.0 Example 92. Synt

Compound 49 was prepared analogously to example 37 (alternate procedure) where 2,2'- {[(benzyloxy)carbonyl]azanediyl}diacetic acid was replaced with N-[(benzyloxy)carbonyl]-L-aspartic acid in the first step of that sequence, lon(s) found by LCMS: (M+3H)/3 = 852.8, (M+4H)/4 = 639.9, (M+5H)/5 = 512.1 , (M+6H)/6 = 427.0

Example 93. Synthesis of Compound 50

Step a. Synthesis of Gly-IN

To a mixture of Z-glycine (92 mg, 0.44 mmol) and HOBT hydrate (94.2 mg, 0.616 mmol) in anhydrous DMF (1 ml) was added EDC-HCI (92.4 mg, 0.484 mmol). The resulting mixture was stirred for 15 minutes, then added with INT-18 (671 .2 mg, 0.4 mmol), followed by DIPEA (390 mg, 3 mmol). After stirred for 3 hours, the reaction mixture was purified through RPLC (50 g, 20 to 90 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (LCMS: [M-2Boc)/2]+ = 778). The material was re-dissolved in MeOH (10 ml) and added with Pd/C. The mixture was stirred under hydrogen overnight. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried under high vacuum. Yield 586.4 mg, 90.4 %. Ion found by LCMS: [(M-Boc)/2]+1 = 760. Step 2. Removal of the Boc group

The reaction was preformed analogously to the removal of the Boc group of deca-Boc Compound 42 precursor. Ions found by LCMS: [M/3]+ = 891 , [M/4]+ = 668, [M/5]+ = 535 , [M/6]+ = 446.

Example 94. Synthesis of Compound 51

The title compound was prepared analogously to Compound 50. Ions found by LCMS: [M/3]+ = 928, [M/4]+ = 696, [M/5]+ = 557 , [M/6]+ = 464, [M/7]+ = 398.

Example 95. Synthesis of Compound 52

The title compound was prepared analogously to Compound 50. Ions found by LCMS: [M/3]+ = 937, [M/4]+ = 703, [M/5]+ = 563, [M/6]+ = 469, [M/7]+ = 402. Example 96. Synthesis of Compound 53

Step a. Synthesis of undeca-Boc-Compound 53

A solution of intermediate INT-20 (0.148 g, 0.0868 mmol), intermediate INT-26 (0.021 Og, 0.0423), and DIEA(0.045ml_, 0.260mmol), in DMF(3ml_), was treated with a solution of HATU(0.0330g,

0.0868mmol) in DMF(1 mL) dropwise via syringe pump over 1 hr. After 1 .5 hr the mixture was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% methanol and water, using no modifier. Yield 0.127 g, 77%. lon(s) found by LCMS: did not ionize

Step b. Synthesis of Compound 53

A solution of undeca-Boc-Compound 53 (0.127 g, 0.0327 mmol), dissolved in DCM (1 mL), was treated with TFA (1 mL), while stirring at room temperature. After 5 minutes, the product was

concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using 0.1 % formic acid as the modifier. Yield 0.090 g, 85% yield, lon(s) found by LCMS: (M+4H)/4 = 692.2, (M+5H)/5 = 553.9, (M+6H)/6 = 461 .8, (M+7H)/7 = 395.9

Example 97. Synthesis of Compound 54a and Compound 54b (Structures assigned arbitrarily)

Procedure A

The title compounds were prepared from INT-38a and INT-38b and INT-18 in a manner similar to that described in Example 73. Yield: 5% for Compound 54a and 51 % for Compound 54b. LC/MS

[M+3H]/3 = 937.4 (for both isomers).

Procedure B (Compond 54a)

Step a. Synthesis of diastereomer a and diastereomer b

HATU (3.1 g, 8.1 mmol) in DMF (5 mL) was added, dropwise, to a solution of racemic-transl -

(ferf-butoxycarbonyl)pyrrolidine-3,4-dicarboxylic acid (1 g, 3.9 mmol), H-Norleu-OMe-hydrochloride (1 .5 g, 8.1 mmol), and triethylamine (4.0 g, 39.5 mmol) in DMF (10 mL) over a period of 20 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 1 0% to 95% acetonitrile and water using 0.1 % TFA modifier. The two diastereomers were separated and pooled and lyophilized separately to yield a more polar compound (diastereomer a) and a less polar compound (diastereomer b) by reversed phase HPLC: LC/MS [m+H]+ = 414.2 (loss of 1 Boc group) for either Boc-protected

intermediates. Each diastereomer was stirred in a 1 /1 mixture of DCM/TFA (8 mL) for 20 minutes then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 0% to 75% acetonitrile and water using no modifier. Yield: 88%, 2 steps. LC/MS [m+H]+ = 414.2.

Note: The configuration of the more polar isomer (diastereomer a) has been arbitrarily assigned the (S,R,R,S) stereochemistry and the configuration of the less polar isomer (diastereomer b) has been arbitrarily assigned the (S,S,S,S) stereochemistry.

Step b. Conjugation of diastereomer a with INT-2

EDC (0.22 g, 1 .16 mmol) was added to a stirring mixture of deprotected diastereomer a (from Step a) (0.40g, 0.97 mmol), 4-[(2-{2-[(6-deoxy-alpha-L mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoic acid (INT-2) (0.41 g, 1 .1 6 mmol), HOBT (0.18g, 1 .1 6 mmol) and triethylamine (0.12 g, 1 .16 mmol) in DMF (4 mL). The mixture was stirred for 2 hours and then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford 660 mg of the di-ester intermediate; LC/MS [m+H]+ = 747.6. The di-ester was stirred in a 1 /1 /2 mixture of methanol/THF/DI water (6 mL) containing LiOH (46 mg, 1 .93 mmol) at ambient temperature for 20 minutes at which point LC/MS analysis confirmed complete hydrolysis to the di-acid product. The mixture was acidified to pH~5 with glacial acetic acid and the volatile solvents were removed by rotovap. The mixture was applied to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were pooled and lyophilized to afford Rhamnose-706-linker A as a white solid. 71 %, 2 steps. LC/MS [m-H]- = 717.2. LC/MS [m+H]+ = 719.2.

Step c. Synthesis of Compound 54a

The title compound was prepared using the product of Step b and INT-18 using a procedure similar to that described in Example 73 for Compound 30. Yield: 53% for Compound 54a LC/MS

[m/3+H]+ = 937.4.

Procedure C (Compound 54b)

Compound 54b was prepared by an analogous procedure as described for Compound 54a in Procedure B, Steps b and c, using diastereomer b and INT-18. Yield 54%. LC/MS [m/3+H]+ = 937.4. Example 98. Synthesis of Compound 55

Step a. Synthesis of L-Arg-INT-18

A mixture of Z-Arg-OH (68 mg, 0.22 mmol) and INT-1 8 (212 mg, 0.2 mmol) was dissolved in anhydrous DMF (1 ml) and DIPEA (91 mg, 0.7 mmol). The solution was cooled in an ice-water bath and added with a solution of HATU (83.6 mg, 0.22 mmol) in DMF (0.5 ml) via syringe pump at a rate of 0.5 ml/hr. After the addition, the mixture was stirred for 30 more minutes, then directly purified through RPLC (50 g, 10 to 90 % acetonitrile and water). The collected fractions were concentrated to a white solid (LCMS: [(M-2Boc)/2]+ = 626). The material was re-dissolved in MeOH (~ 10 ml), and the solution was added with Pd/C. The resulting mixture was stirred under hydrogen for 4 hours. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried in high vacuum to afford the title compound as a white solid (236.8 mg, 97.2 %). Ion found by LCMS: [M/2]+ = 610.

Step b. Synthesis of Octa-Boc Compound 55 precursor

A mixture of INT-40 (40 mg, 0.0347 mmol), L-Arg-INT-18 (90 mg, 0.07 mmol) was dissolved in in anhydrous DMF (0.5 ml) and DIPEA (52 mg, 0.4 mmol). The solution was cooled in an ice-water bath and added with a solution of HATU (26.6 mg, 0.07 mmol) in DMF (0.5 ml) via syringe pump at a rate of 0.5 ml/hr. After the addition, the reaction was stirred for 30 more minutes. It was then purified through RPLC (50 g, 20 to 100 % MeOH and water). Yield 72 mg, 53.7 %. Ions found by LCMS: (M+3H)/3 = 1251 , [(M- Boc)/3]+= 1218, [(M-5Boc)/3]+= 1084.

Step c. Removal of the Boc groups

Compound 55 Boc intermediate (72 mg, 0.0186 mmol) was dissolved in TFA (~ 0.5 ml). The solution was stirred at room temperature for 15 minutes, then directly purified through RPLC (50 g, 5 to 38 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 27 mg, 35.5 %. Ions found by LCMS: [M/3]+ = 984, [M/4]+ = 738, [M/5]+ = 591 , [M/6]+ = 492, [M/7]+ = 422.

Example 99. Synthesis of Compound 56

The title compound was prepared analogously to Compound 55. Ions found by LCMS: [M/3]+ = 956, [M/4]+ = 717, [M/5]+ = 574, [M/6]+ = 478, [M/7]+ = 410. Example 100. Synthesis of Compound 57

The title compound was prepared analogously to Compound 55. Ions found by LCMS: [M/3]+ 965, [M/4]+ = 724, [M/5]+ = 580, [M/6]+ = 483.

Example 101. Synthesis of Compound 58

The title compound was prepared analogously to Compound 55. Ions found by LCMS: [M/3]+ 972, [M/4]+ = 730, [M/5]+ = 584, [M/6]+ = 487.

Example 102. Synthesis of Compound 59

Step a. Preparation of di-tert-butyl N,N-bis[2-(benzyloxy)-2-oxoethyl]-L-aspartate

Benzyl Bromo-acetate (4.6 g, 20 mmol) was added into the solution di-tert-butyl L-aspartate (2.45 g, 10 mmol) and DIPEA (3.6 mL, 30 mmol) in 60 mL DMF, the resultant solution was stirred at room temperature overnight. The reaction solution was concentrated and purified by flash chromatography to provide products. Yield of oil product, LC/MS, 542.3 [M+H]+.

Step b. Preparation of N,N-bis[2-(benzyloxy)-2-oxoethyl]-L-aspartic acid

The above product di-tert-butyl N,N-bis[2-(benzyloxy)-2-oxoethyl]-L-aspartate (1 g, 1 .84 mmol) was treated with 5 mL trifluoroacetic acid and 5 mL DCM, the solution was stirred for a half hour and then concentrated and dried under high vacuum used for next step without purification. Yield of oil product, LC/MS, 430.1 [M+H]+.

Step c. Preparation of dibenzyl 2,2'-{[(2S)-1 ,4-dioxo-1 ,4-bis{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5- trihydroxy-6-methyloxan-2-yl]oxy}ethoxy)ethyl]amino}butan-2-yl]azanediyl}diacetate

The above crude product N,N-bis[2-(benzyloxy)-2-oxoethyl]-L-aspartic acid (1 .84 mmol) was redissolved in 20 mL DMF, then INT-1 . (1 .25 g, 5 mmol), EDC (1 .0 g, 5 mmol), HOBT (0.75 g, 5 mmol), Hunig's base (1 .4 mL, 10 mmol) were added the solution at room temperature. The resultant solution was stirred overnight. The resultant solution was removed DMF and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. LC/MS 896.4 [M+H].

Step d. Preparation of 2,2'-{[(2S)-1 ,4-dioxo-1 ,4-bis{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyloxan-2-yl]oxy}ethoxy)ethyl]amino}butan-2-yl]azanediyl}diacetic acid Dibenzyl 2,2'-{[(2S)-1 ,4-dioxo-1 ,4-bis{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2- yl]oxy}ethoxy)ethyl]amino}butan-2-yl]azanediyl}diacetate (500 mg, 0.56 mmol) was dissolved into 5 mL MeOH and 5 mL ethyl acetate, then 200 mg of 5% palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere overnight. The palladium charcoal was removed by filtration after completed the reaction by LCMS. The filtrate was concentrated and used next step without any purification. LC/MS, 71 6.3 [M+H]+ .

Step e. Preparation of per-Boc-Compound 59 (Standard Procedure A)

To the solution of 2,2'-{[(2S)-1 ,4-dioxo-1 ,4-bis{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyloxan-2-yl]oxy}ethoxy)ethyl]amino}butan-2-yl]azanediyl}diacetic acid (72 mg, 0.1 mmol), triethyl amine (0.07 mL, 0.5 mmol) and INT-20 (340 mg, 0.2 mmol) in 5 mL DMF was added HATU (78 mg, 0.2 mmol). The reaction was stirred for 1 hr and then the resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of product (per-Boc-Compound 59), 220 mg, 65% yield.

Step f. Preparation of Compound 59 (Standard Procedure B)

Per-Boc-Compound 59 from previous step was treated in 2 mL DCM and 2 mL trifluoroacetic acid at room temperature, the solution was stirred 10 min, then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using trifluoroacetic acid as the modifier. Yield 0.120 g, 67% yield, lon(s) found by LCMS: [M/3]+1 = 1029.9, [M/4]+1 =772.7, [M/5]+1 =61 8.4, [M/6]+1 =515.5.

Example 103. Binding of Selected Compounds from Examples 23-49, 69-102, and 163-165 to Lipopolysaccharide (LPS)

Binding of compounds to LPS derived from Pseudomonas aeruginosa (ATCC 27316) was determined according to the following procedure. LPS from other strains of bacteria may be substituted to determine binding of compounds with any particular LPS sample.

Step a. Synthesis of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-

A solution of methyl (2S)-2-[(N-{(2S)-2-amino-4-[(tert-butoxycarbonyl)amino]butanoyl}-L- threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.600 g, 1 .124 mmol),(2S)-2- {[(benzyloxy)carbonyl]amino}octanoic acid (0.330 g, 1 .124 mmol), and DIEA (0.59 mL, 3.37 mmol) in DMF(4 mL), was treated with HATU (0.470 g, 1 .24 mmol), and stirred for 30min. The reaction was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 10% to 100% acetonitrile and water, using formic acid (0.1 %) as the modifier. Yield 0.726 g, 79% yield, lon(s) found by LCMS: (M+H)⁺ = 809.5

A mixture of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-{[(benzyloxy)carbonyl]amino}octanoyl]amino}-4- [(tert-butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.300 g, 0.371 mmol), 5%Pd/C(0.150g), and methanol(4 mL), were charged with hydrogen from a balloon. After 2hr the product was filtered through Celite, concentrated, and used in the next step without further purification. Mass recovery 0.250 g, quantitative, lon(s) found by LCMS: (M+H)⁺ = 675.4

Step c. Synthesis of methyl (2S)-4-[(tert-butoxycarbonyl)amino]-2-({N-[(2S)-4-[(tert- butoxycarbonyl)amino]-2-{[(2S)-2-{[5-(dimethylamino)naphthalene-1- sulfonyl]amino}octanoyl]amino}butanoyl]-L-threonyl}amino)butanoate

A solution of methyl (2S)-2-[(N-{(2S)-2-{[(2S)-2-aminooctanoyl]amino}-4-[(tert- butoxycarbonyl)amino]butanoyl}-L-threonyl)amino]-4-[(tert-butoxycarbonyl)amino]butanoate (0.60 g), and dansyl chloride (0.1 10 g, 0.408 mmol), in NMP (1 .5 mL) was treated with N-methylmorpholine (0.081 mL, 0.741 mmol). After 30 minutes the product was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% acetonitrile and water, using formic acid (0.1 %) as the modifier. Yield 0.336 g, 85% yield, lon(s) found by LCMS: (M+H)⁺ = 908.5

Step d. Synthesis of (2S)-4-[(tert-butoxycarbonyl)amino]-2-({N-[(2S)-4-[(tert- butoxycarbonyl)amino]-2-{[(2S)-2-{[5-(dimethylamino)naphthalene-1- sulfonyl]amino}octanoyl]amino}butanoyl]-L-threonyl}amino)butanoic acid

A solution of methyl (2S)-4-[(tert-butoxycarbonyl)amino]-2-({N-[(2S)-4-[(tert- butoxycarbonyl)amino]-2-{[(2S)-2-{[5-(dimethylamino)naphthalene-1 - sulfonyl]amino}octanoyl]amino}butanoyl]-L-threonyl}amino)butanoate (0.285 g, 0.314 mmol), in methanol (10 mL), was treated with lithium hydroxide (0.050 g, 2.1 mmol) in water(1 mL), while stirring at room temperature. After 30 minutes the mixture was titrated with concentrated HCI(aq.) until slightly acidic by pH paper, concentrated, and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using no modifier. Yield 0.267 g, 95% yield, lon(s) found by LCMS: LCMS: (M+H)⁺ = 894.5

Step e. Synthesis of penta-Boc-(Compound 60)

A solution of (2S)-4-[(tert-butoxycarbonyl)amino]-2-({N-[(2S)-4-[(tert-butoxycarbonyl)amino]-2-

{[(2S)-2-{[5-(dimethylamino)naphthalene-1 -sulfonyl]amino}octanoyl]amino}butanoyl]-L- threonyl}amino)butanoic acid(0.267 g, 0.299 mmol), INT-1 1 (0.349 g, 0.328 mmol), and DIEA (0.172 mL, 0.985 mmol) in DMF (1 mL), was treated with HATU (0.129 g, 0.328 mmol), while stirring at room temperature. The product was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% methanol and water, using no modifier. Yield 0.267 g, 46% yield, lon(s) found by LCMS: (M+2H)/2 = 969.5, [(M-1 Boc)+2H]/2 = 919.5, [(M-2Boc)+2H]/2 = 869.5

Penta-Boc-(Compound 60) (0.267 mg, 0.138 mmol) was treated with TFA (3 mL) for 15 minutes, then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco

CombiFlash liquid chromatograph eluted with 0% to 1 00% acetonitrile and water, using formic acid as the modifier. The appropriate fractions were collected, pooled and concentrated to give after drying and lyophilization, Compound 60. Yield 0.053 g, 27% yield, lon(s) found by LCMS: [M+2H]/2 = 719.4 Part 2. Binding assay

The LPS binding of compounds was measured by fluorescence of a compound bound to LPS using a Tecan Infinite Pro fluorescence spectrophotometer set at an excitation wavelength of 340 nm and emission wavelength of 485 nm. Displacement assays were performed by recording the fluorescence after the addition of inhibitors at 250 μΜ final concentration. Assays were performed in 96-well microtiter plates using 4 μg of P. aeruginosa LPS (ATCC 27316) and 6 μΜ Compound 60 in 10 mM HEPES buffer (pH 7.5). Percent displacement is expressed relative to a no inhibitor control and corrected for intrinsic inhibitor fluorescence.

Part 3. Results

Table 3. % Displacement of Fluorescent Probe Compound 60 from P. aeruginosa LPS

Compound % Displacement Compound % Displacement

Compound 29 97.3 Compound 58 49.2

Compound 30 79.7 Compound 59 94.2

Compound 31 96 Compound 75 90.1

Compound 32 97 Compound 76 93.1

Compound 33 98.8 Compound 77 87.5

Compound 42 92.2 Compound 78 86.9

Compound 43 87.9 Compound 79 95.7

Compound 44 48.5 Compound 80 97.6

Compound 45 58.6

Example 104. Antibacterial Activity of Selected Compounds from Examples 23-49, 69-101 , and 151-209

Part 1. Methods

Generation of E. co/ BW25113 + pUC18-mcr-1.

An mcr- 1 expression plasmid was constructed (GenScript; Piscataway, NJ) using the pUC18 high copy number plasmid (Yanisch-Perron, et al., 1985). A 1649-bp fragment was synthesized containing, in order from 5' to 3': EcoR1 restriction site (5'-GAATTC-3'), a ribosomal binding site (5'-AGGAGG-3'), a 5- bp native start codon upstream sequence (5'-TTCTC-3') from the mcr- /-bearing plasmid pHNSHP45

(Liu, YY, et al., 2015; GenBank Accession # KP347127), the 1626-bp mcr- 1 open reading frame with stop codon ("orf00073" of GenBank Accession # KP347127), and finally an Xba\ restriction site sequence (5'- TCTAGA-3'). Restriction digestion and subsequent ligation into the multiple cloning site of the pUC18 backbone resulted in directional integration of the mcr- 1 gene. The pUC1 8-mcr-1 plasmid was transformed into chemically-competent E. coli BW251 13 (Coli Genetic Stock Center #7636) as described previously (Chung, et al., 1989), recovered for 1 h shaking at 37°C in Super Optimal broth with Catabolite repression (SOC) media and then aliquots were plated on Luria-Bertani (LB) media containing 100 μg/mL of carbenicillin. Putative transformant colonies were then verified in MIC assays. Strains and culture conditions.

Bacterial strains were stored as glycerol stocks at -80 °C prior to culturing on cation-adjusted

Mueller-Hinton agar (MHA; BD cat. no. 21 1438) or in cation-adjusted Mueller-Hinton broth (MHB; BD cat. no.212322) cultured at 37°C. Antibacterial activity was assessed against a strain panel consisting of

Escherichia coli BW251 13 wild-type (Coli Genetic Stock Center #7636), BW251 13 + pUC1 8-mcr-1 , BW251 13 COL^r 4X-12 mutant (an uncharacterized, spontaneous mutant selected by plating BW251 13 on

MHA containing 1 μg/mL COL, or 4X the MIC value), and ATCC BAA-2469 (ATCC - American Type

Culture Collection), Acinetobacter baumannii ATCC A5075 (ATCC), Pseudomonas aeruginosa PA01

(ATCC BAA-47, ATCC), and Klebsiella pneumoniae ATCC 10031 (ATCC). MIC (Minimum Inhibitory Concentration) assays.

MIC assays were performed according to CLSI broth microdilution guidelines (M07-A9, M100- S23) with the exception of using a 1 00 μΙ_ assay volume and preparing stock compounds at 50X final concentration. Briefly, stock solutions of all antibacterial agents were prepared fresh in appropriate solvents (i.e. DMSO, Dl water, ethanol, etc.). Stock concentrations were made at 50X the highest final assay concentration and serially diluted 2-fold, 12 times in a 96-well PCR plate (VWR cat. no. 83007- 374). Bacterial cell suspensions generated from MHA plate cultures were prepared in 0.85% saline and adjusted to -0.1 Οϋβοο nm. Next, cell suspensions were diluted 1 :200 in MHB (BD cat. no. 212322) to a concentration of ~5 x 10⁵ CFU (Colony Forming Units)/ml_. Ninety-eight microliters of each cell suspension in MHB were added to test wells in a 96-well assay plate (Costar cat. no. 3370). A Beckman Multimek 96 liquid handling robot was used to dispense 2 μΙ_ of each 50X stock compound into the plate containing 98 μΙ_ of each strain in MHB (2% final solvent concentration). Plates were shaken then incubated at 37°C overnight (16-20 h). MIC values were read visually at 100% growth inhibition for all compounds.

Part 2. Results

Table 4. MICs ^g/mL)

^* ND: not determined

Example 105. Synergistic Activity of Selected Compounds from Examples 23-35, 38-49, 69-101 , and 151-172

Fixed concentration synergy MIC assays. The synergistic potential of test compounds was investigated through assessment of MIC values in the presence of a fixed concentration of erythromycin (ERY; Sigma cat. no. E5389). The test compound was run as a dilution series as performed under the CLSI-based conditions described in Example 51 for single agents using E. coli BW251 13 (or E. coli BW251 13 + pUC18-mcr-1 ), however each well was stamped a subsequent time with Beckman Multimek 96 liquid handling robot to aliquot a 2 μΙ_ volume of 0.05 mg/mL ERY (solubilized in 50% ethanol) across the 12-point, 2-fold dilution series to give a resulting concentration of ~1 μg/mL ERY. MIC values for the test compounds alone were compared with those containing the fixed concentration of ERY to determine if the polymyxin-based compound increased the susceptibility of the strain to ERY. Table 5. MICs in combination/absence of 1 μg/mL erythromycin and resultant fold-synergy

E. coli + 1 £. co/ + 1

Fold Fold

Compound E. coli Mg/mL E. coli Mg/mL

Synergy Synergy erythromycin erythromycin

BW25113

BW25113 +

BW25113 BW25113 + pUC18- pUC18-mcr-1 mcr-1

Compound 22 16 1 16 256 4 64

Compound 23 128 2 64 256 128 2

Compound 24 2 1 2 8 2 4

Compound 25 2 2 1 4 4 1

Compound 26 4 2 2 16 16 1

Compound 27 1 1 1 1 1 1

Compound 28 2 2 1 2 2 1

Compound 29 4 4 1 4 4 1

Compound 30 1 1 1 128 2 64

Compound 31 4 2 2 4 2 2

Compound 32 2 2 1 2 2 1

Compound 33 2 1 2 2 2 1

Compound 34 2 2 1 256 256 1

Compound 35 1 1 1 4 2 2

Compound 36 0.5 0.5 1 32 16 2

Compound 37 1 1 1 32 32 1

Compound 38 1 1 1 16 16 1

Compound 39 2 2 1 2 2 1

Compound 40 4 4 1 8 4 2

Compound 41 4 2 2 4 2 2

Compound 42 2 2 1 4 2 2

Compound 43 1 2 0.5 64 64 1

Compound 44 2 2 1 4 4 1

Compound 45 4 2 2 8 4 2

Compound 46 2 2 1 2 2 1

Compound 47 2 2 1 2 2 1 £. coli + 1 E. coli + 1

Fold Fold

Compound E. coli Mg/mL £. co/ Mg/mL

Synergy Synergy erythromycin erythromycin

BW25113

BW25113 +

BW25113 BW25113 + pUC18- pUC18-mcr-1 mcr-1

Compound 48 1 1 1 16 64 0.25

Compound 49 2 1 2 16 16 ¹

Compound 50 2 1 2 64 64 ¹

Compound 51 2 1 2 8 2 4

Compound 52 2 2 2 2

Compound 53 2 2 ¹ 2 2 ¹

Compound 54a 1 1 ¹ 1 2 0.5

Compound 54b 2 1 2 2 2 ¹

Compound 55 2 2 ¹ 4 4 ¹

Compound 56 2 2 ¹ 1 2 0.5

Compound 57 2 2 ¹ 2 2 ¹

Compound 58 4 4 ¹ 8 8 ¹

Compound 59 2 2 1 2 2 1

Compound 61 1 1 1 64 64 1

Compound 62 1 1 1 32 32 1

Compound 63 2 1 2 128 64 2

Compound 64 4 2 2 4 4 1

Compound 65 2 4 0.5 2 4 0.5

Compound 66 2 1 2 2 2 1

Compound 67 2 2 1 4 2 2

Compound 68 2 2 1 2 2 1

Compound 69 1 1 1 2 2 1

Compound 70 2 2 1 2 2 1

Compound 71 1 1 1 64 64 1

Compound 72 2 2 1 2 2 1

Compound 73 1 1 1 2 2 1

Compound 74 2 2 1 2 4 0.5

Compound 75 4 2 2 16 4 4

Compound 76 4 4 1 8 4 2 E. coli + 1 £. co// + 1

Fold Fold

Compound E. coli Mg/mL £. co/ Mg/mL

Synergy Synergy erythromycin erythromycin

BW25113

BW25113 +

BW25113 BW25113 + pUC18- pUC18-mcr-1

mcr-1

Compound 77 4 4 1 32 4 8

Compound 78 4 4 1 8 4 2

Compound 79 4 2 2 2 2 1

Compound 80 4 4 1 4 4 1

Compound 81 2 2 1 2 2 1

Compound 82 2 2 1 2 2 1

Compound 83 4 4 1 4 4 1

Compound 84 1 1 1 1 1 1

Compound 85 2 2 1 2 2 1

Compound 86 2 1 2 2 1 2

Compound 87 1 1 1 1 1 1

Example 106. Synergistic Activity of Compound 1

Checkerboard synergy assays.

Checkerboard synergy assays were performed as previously described (Eliopoulos, G.M. and R.C. Moellering, 1996; Moody, J., 2004). In master stock plates the stocks of two compounds

("Compound A" - test compound, and "Compound B" - a known antibacterial agent) for synergistic evaluation were prepared. A liquid stock of Compound A was prepared fresh in Dl water to a concentration 50X the desired final synergy assay concentration. Into wells A1 through H1 (vertical, Y- axis column on left edge of plate) of a 96-well PCR plate (VWR cat. no. 83007-374) Compound A was aliquoted in 100 μΙ_ volumes. To all other wells on the plate, with the exception of A12 through H12, 50 μΙ_ of Dl water was added. A multichannel pipette was then used to transfer 50 μΙ_ from column A1 -H1 to column A2-H2. Following mixing, tips were discarded and the process was repeated across the plate to create a series of 1 1 , serial two-fold dilutions spanning columns 1 through 1 1 .

A stock of Compound B was prepared in a similar fashion in the appropriate solvent (DMSO, Dl water, ethanol, etc.) at 50X the desired final synergy assay concentration. In a separate 96-well PCR plate, 100 μΙ_ aliquots of Compound B were added to wells in the top row of the plate spanning A1 through A12. In all other wells, excluding the bottom row (H1 -H12), 50 μΙ_ of the corresponding Compound B solvent were added. In a similar fashion to Compound A, Compound B was serially diluted, two-fold down the plate using a multichannel pipette, resulting in 7 rows comprised of 12 wells with 50 μΙ_ diluted Compound B. Checkerboard synergy assays were performed using the same assay setup as the standard MIC assays previously described previously herein (i.e. inoculum, media, volume, 96-well assay plates, incubation time, incubation temperature, 100% inhibition assay endpoint, etc.). However, rather than a single addition of 2 μΙ_ of 50X stock compound solution to the assay plate containing MHB pre-inoculated with bacterial cells using the Multimek 96 liquid handling robot, two sequential additions were made (one from the Compound A 50X stock plate and one from the Compound B 50X stock plate). The resulting checkerboard plates contained a 7 x 1 1 -well grid (A1 -G1 x A1 -A1 1 ) of synergy wells containing varying proportions of Compounds A and B, an 1 1 -point dilution series of Compound A alone (wells H1 -H1 1 , assay concentration range ^"I X to 1 /1024X), a 7-point dilution series of Compound B alone (wells A12- G12, assay concentration range ^"I X to 1 /64X) and a positive growth control well containing no drug (well H12) (FIG. 1 ).

Following overnight incubation, checkerboard synergy plates were read visually at 100% growth inhibition. MIC values for Compound A and B were recorded from their respective single-drug dilution series wells and an optimal synergy well (or wells) was identified as that for which growth inhibition was achieved using a combination of the lowest possible concentrations of both drugs. The fraction inhibitory concentration (FIC) was calculated for each compound by dividing the combination MIC value in the optimal synergy well by the MIC value for the compound alone. The summation of the FIC values for Compounds A and B was then used to calculate the fractional inhibitory concentration index (FIG) and synergy was defined as an FICI value of <0.50.

FIC Compound A = MIC of Compound A in combination/MIC of Compound A alone

FIC Compound B = MIC of Compound B in combination/MIC of compound B alone

Table 6. MICs of Compound 1 (CMP 1 ) and various antibiotics against Gram-negative bacterial strains and FICI values of Compound 1 in combination with those antibiotics

Example 107. Inoculation of mice or rabbits with OVA-Rha-linked vaccine, isolation of serum and purification of anti-Rha antibodies

Part 1. Preparation of OVA-Rha-linked vaccine (OVA-Rha)

Step a. Synthesis of N-(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)-4-[(2,5- dioxopyrrolidin-1-yl)oxy]-4-oxobutanamide

(0-(N-Succinimidyl)-1 ,1 ,3,3-tetramethyl uronium tetrafluoroborate-' STU") (206 mg, 0.68 mmol) was added to a stirring mixture of 4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanoic acid (INT-2) (200 mg, 0.57 mmol) and triethylamine (690 mg, 0.68 mmol) in DMF (4 mL) and the reaction was stirred for 4 hours at room temperature. The solvent was removed on a rotary evaporator then dried under high vacuum for 12 hours. The crude product mixture was used for protein conjugation without further treatment. (~2 mg of the crude product was treated with excess benzylamine and analyzed by LC/MS to confirm formation of the activated ester). Step b. Conjugation to ovalbumin

A solution of N-(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)-4-[(2,5-dioxopyrrolidin- 1 -yl)oxy]-4-oxobutanamide (30 mg/mL in 2x PBS pH 7.4) was added to the same volume of ovalbumin (10 mg/mL in 2x PBS pH 7.4) and was gently stirred at 25°C for 1 h, followed by 5 h incubation at 4°C. The solution was concentrated and buffer exchanged into 1 x PBS pH 7.4 using Amicon Centrifugal Filter Devices (10,000 MWCO). Extensive buffer washes removed unconjugated linker from the protein solution. Solution purity and a molecular weight shift were confirmed by SDS-PAGE and size exclusion chromatography (Superdex 75). The solution was determined to be endotoxin free using the Endosafe Assay (Charles River). The glycoprotein solution was quantitated using a NanoDrop 2000

Spectrophotometer and stored at 4°C for immunizations.

Part 2. Vaccination of animals for production of anti-Rha serum

Mice were vaccinated subcutaneously in a similar manner to that described in ACS Chem Biol. 201 1 Feb 18;6(2):1 85-91 . Female ICR mice (17-20 g each) were immunized q2wksx4 with 30 μg of OVA-Rha as follows: Day 0: each mouse received a subcutaneous injection of 150 μΙ_ of OVA-Rha + Complete Freund's Adjuvant (CFA, Sigma F5881 ); Day 14: 1 50 μΙ_ of OVA-Rha + Incomplete Freund's Adjuvant (IFA Sigma F5506); Day 28: 150 μΙ_ OVA-Rha + saline. An additional immunization on Day 42 using 150 μΙ_ OVA-Rha + saline can be used to further increase titers of anti-Rha antibody. Serum was isolated subsequently from mouse blood according to standard protocol. Serum production may be accomplished at least 7 to 21 days after the last vaccination. Part 3. Vaccination of rabbits

Sixty day old New Zealand rabbits were vaccinated subcutaneously with OVA-Rha ± CFA or IFA. Rabbits were held at indicated time points and serum wasw prepared using a standard procedure.

Week 0: 1 st immunization with 0.25 mg of OVA-Rha in Complete Freund's Adjuvant

Week 2: 2nd immunization with 0.2 mg of OVA-Rha in Incomplete Freund's Adjuvant (IFA)

Week 4: 3rd immunization with 0.2 mg of OVA-Rha in IFA

Week 5: 1 st production bleed - -20-25 mL of serum

Week 6: 4th immunization with 0.2 mg of OVA-Rha in IFA

Week 7: 2nd Production bleed - -20-25 mL of serum

Week 8: 3rd Production bleed - -20-25 mL of serum

Part 4. Purification of Rha-specific antibodies from serum

Step a. Preparation of the anti-rhamnose antibody affinity column

6-Aminohexyl-Agarose (5 mL, 4% crosslinked beaded agarose, -5 μιηοΙ per mL) suspended in saline was centrifuged (3500 rpm for 5 min), then decanted to remove water. The resulting solid was suspended in DMF (4 x 15 mL), centrifuged (3500 rpm for 5 min) and decanted. The agarose was resuspended in DMF (1 mL), and treated with INT-2 (0.140 g, 0.40 mmol), followed by DIEA (0.21 mL, 1 .20 mmol), and HATU (0.152 g, 0.40 mmol). The mixture was rotated for 3 hr at room temperature, then transferred to a plastic column and washed with phosphate buffered saline (5 x 15 mL, pH 7.4). The solid supported a-L-rhamnose was stored in phosphate buffered saline (pH 7.4) with 20% ethanol, at room temperature.

Step b. Purification of anti-rhamnose antibody from rabbit serum

Serum from immunized rabbits was diluted 2-fold in 2X PBS pH 7.4. The solution was passaged over the solid supported a-L-rhamnose column (from Step a) by gravity flow followed by a wash step using 10 CV of 1 X PBS pH 7.4. Protein was eluted using 20 mM Glycine Buffer pH 2.0 and immediately adjusted to a neutral pH by the addition of 1 M Tris pH 9.0. The eluate was buffer exchanged with 1 X PBS pH 7.4 and concentrated in Amicon centrifugal concentrators (10K MWCO). The presence of IgG was determined by reducing and non-reducing SDS-PAGE and quantitated using a NanoDrop 2000 Spectrophotometer. The resultant purified antibodies can be used in place of serum adjusting for relative antibody titers.

Example 108. Rha binding assay - Plate ELISA

Part 1. Binding of anti-rhamnose (anti-Rha) antibodies to selected compounds from Examples 23- 49 and 69-101

Test compound (2 μg) was immobilized to 96 well microtiter plates using the TaKaDa peptide coating kit (Clontech). Binding was detected with 3-fold dilutions of polyclonal rabbit anti-rhamnose serum (from Example 54) or na^'ive rabbit serum (as a control), starting with a 1 :450 dilution, and ending with a 1 :984,150 dilution. Bound rabbit antibody was detected using a 1 :5,000 dilution of anti-rabbit IgG HRP antibody and TMB development solution. After stopping the reaction with 1 M H2SO4, the absorbance at 450 nm (A450 nm) was read using a Magellan plate reader and plotted against serum dilution factor using GraphPad Prism and Microsoft Excel for data analysis. A BCso was calculated and is the reciprocal of the serum dilution at 50% maximum absorbance at 450 nm. Part 2. Results

Table 7. BCso values for selected Compounds from Examples 23-49 and 69-101

^* Reciprocal of serum dilution giving 50% signal

Example 109. Rha binding assay - Whole cell ELISA

Part 1. Binding of anti-rhamnose antibodies to selected E. co//^'-bound compounds 23-49 and 69- 101.

96 Well microtiter plates coated with E. coli were incubated with a 5-fold dilution series of test compound, starting with 4,000 ng and ending with 0.0512 ng. Binding was detected with a fixed

1 :100,000 dilution of polyclonal rabbit anti-rhamnose serum (from Example 54) or naive rabbit serum (as a control). Bound rabbit antibody was detected using a 1 :5,000 dilution of anti-rabbit IgG HRP antibody and TMB development solution. After stopping the reaction with 1 M H2SO4, the absorbance at 450 nm (A450 nm) was read using a Magellan plate reader and plotted against the amount of compound in ng using GraphPad Prism and Microsoft Excel for data analysis. Part 2. Results

Table 8 below shows the ECso values for selected Compounds from Examples 23-49 and 69-101 . FIG. 15 shows the absorbance values at 450 nm for E. coli incubated with Compound 14 and a control compound (Compound 14 without the monosaccharide portion).

Table 8

^* Concentration of compound that gave 50% of total signal (ng)

Example 110. LPS Neutralization by Selected Compounds from Examples 23-49 and 69-101 Part a. Method

The murine macrophage cell line RAW 264.7 (ATCC, TIB-71 ) was grown to about 70 % confluency in DMEM + 10 % FBS in T75 cm² flasks at 37 °C / 5 % CO2 for 2-3 days. Cells were washed once with 10 mL DPBS and scraped into 5 mL of media. After centrifugation at 800 rpm for 5 min, the cells were counted with a hemocytometer after 1 :10 dilution in media with 0.04 % trypan blue. Cells were adjusted to 2E6/ mL in DMEM + 1 0 % FBS.

RAW 264.7 cells were added to a 96-well plate at 2 x 10⁵ cells per well in 100 μΙ_ in DMEM + 10 % FBS and incubated for 24 h at 37°C 1 5% CO2. Medium was aspirated and cells washed once with 200 μΙ_ DPBS before addition of test reagents. Test reagents and media only control were prepared in RPMI without phenol red + 5% FBS.

Table 9. Test article preparation

Test Compounds Polymyxin B LPS (positive control)

5 \iqlmL· 5 pQlmL 10 \iqlmL·

7.5 μΙ_ 15 μΙ_ Polymyxin B 1 μΙ_ LPS

(2 mg/mL ) (1 mg/mL ) (1 mg/mL )

1492.5 μΐ media 1485 μΐ media 999 μΐ media

1 .5 mL 1 .5 mL 1 mL

100 it of reagents were added to wells at 0.5 and 5 μς/ mL final concentrations. The cells were incubated with reagents for 1 h at 37°C + 5% CO2. After 15 min, 100 μί medium containing LPS at 0.001 , 0.01 , 0.1 , 1 and 10 μς/ mL was added to appropriate wells and the plates were incubated for 24 h at 37°C + 5% CO2. Controls received medium only. After 24h, 150 μΙ of each supernatant was transferred to a new 96 well FLAT bottom plate. 50 μί of the transferred supernatant was used for a Griess assay (measurement of nitric oxide (NO) production). The remainder was stored at 4 °C for ELISA. A Griess assay was performed according to manufacturer's (Promega) instructions. Absorbance at 540 nm was read and NO production in μΜ was calculated by linear regression analysis of the NO standard curve using Graphpad Prism 6 software.

Part b. Results: Inhibition of Nitric Oxide (NO) production by Selected Compounds from

Examples 23-49 and 69-101 in RAW cells stimulated with LPS

NO production in response to LPS (endotoxin) is a signal of macrophage activation which may lead to sepsis in an infected host. Inhibition of NO production through LPS binding and neutralization is demonstrated by Compound 14 of the present disclosure (FIG. 2) at clinically relevant concentrations of LPS that have been correlated with sepsis in humans which is less than 1 ng/mL in the plasma (see, e.g., Opal et al., J. Infect Dis. 180:1584-9, 1999).

Example 111. In vivo activity in a mouse model of bacterial sepsis for selected compounds from Examples 23-49 and 69-101

Mouse strain

Experiments utilized female C57BI/6 mice (18 - 22 grams) from Charles River Laboratories

(Wilmington, Massachusetts) and were allowed to acclimate for 5 days prior to start of study. Animals were housed 6 per cage with access to food and water ad libitum.

Inoculum preparation and infection

E. coli ATCC 25922 was obtained from the American Type Culture Collection (Manassas, Va) and was prepared for infection from an overnight plate culture (Trypicase Soy agar media). A portion of the plate was re-suspended in sterile saline and adjusted to an OD of 0.1 at 625 nm. The adjusted bacterial suspension was further diluted in 5% hog gastric mucin to target an infecting inoculum of 1 .0x10³ CFU/mouse. Plate counts of the inoculum were performed to confirm inoculum concentration. To initiate the infection, mice were injected with 0.5 mL of the suspension intraperitoneally (IP).

Treatment/Efficacy

When indicated, groups of mice received an IV injection of rabbit anti-serum or purified antibody (+Ab) 24 hours prior to infection at a dose of 1 :4000. Mice that received no serum or antibody are indicated by (-Ab). Beginning at one hour post infection, mice were dosed with either vehicle or test article. Vehicle/diluent was sterile 0.9% saline for all test articles. All animals were dosed IP at 10 mL /kg. Mice were euthanized at 16 hours post infection. Kidneys were aseptically removed, weighed, homogenized, serially diluted and plated. The CFUs were enumerated after overnight incubation. The average and standard deviations for each group were determined via accepted methodology. Results

Table 10. Survival and CFU reduction in kidneys of mice (n=6 per group) infected with E. co//^' and treated with colistin or Compound

Table 1 1 . Survival and CFU reduction in kidneys of mice (n=6 or 10 per group) infected with E. co//^' and treated with meropenem or Compound

Table 12. Survival and CFU reduction in kidneys of mice (n=6 per group) infected with E. co//^' and treated with meropenem or Compound

Change in logi₀

Dose (mg/kg) Number of logioCFU/

Group ID St. Dev. CFU/g kidney versus

(IP, QD) survivors g kidney

16 hour controls

5.0 (-Ab) 6 3.30 0.69 -3.78

Compound 1 1

5.0 (+Ab) 6 2.1 1 0.00 -4.97

Additional efficacy was seen in the presence of serum with Compound 1 , Compound 1 1 , and Compound 14 indicating enhanced efficacy due to conjugation of antigen and recognition by anti-Rha antibody. Example 112. Antibacterial Activity of Compounds 14 and 54a

Part 1. Methods

Generation of E. co/ BW25113 + pUC18-mcr-2.

An mcr-2 expression plasmid was constructed (GenScript; Piscataway, NJ) using the pUC18 high copy number plasmid (Yanisch-Perron, et al., 1985). A 1640-bp fragment was synthesized containing, in order from 5' to 3': EcoR1 restriction site (5'-GAATTC-3'), a ribosomal binding site (5'-AGGAGG-3'), a 5- bp native start codon upstream sequence (5'-TTTCT-3') from the mcr-2-bearing plasmid pKP37-BE (Xavier et al., Euro Surveill A (27), 201 6; GenBank Accession # LT598652), the 1 617-bp mcr-2 open reading frame with stop codon (locus tag # A7J1 1_03753), and finally an Xba\ restriction site sequence (5'-TCTAGA-3'). Restriction digestion and subsequent ligation into the multiple cloning site of the pUC18 backbone resulted in directional integration of the mcr-2 gene. The pUC1 8-mcr-2 plasmid was transformed into chemically-competent E. coli BW251 13 (Coli Genetic Stock Center #7636) as described previously (Chung, et al., 1989), recovered for 1 h shaking at 37°C in Super Optimal broth with Catabolite repression (SOC) media and then aliquots were plated on Luria-Bertani (LB) media containing 100 μg/mL of carbenicillin. Putative transformant colonies were then verified in MIC assays. Susceptibility of Compounds 14 and 54a, COL (colistin), and PMB (polymyxin B) were then assessed vs. WT E. coli BW251 13 and the pUC18-mcr-2 transformant strain.

Strains and culture conditions.

Bacterial strains were stored as glycerol stocks at -80 °C prior to culturing at 37°C on cation- adjusted Mueller-Hinton agar (MHA; BD cat. no. 21 1438) or in cation-adjusted Mueller-Hinton broth

(MHB; BD cat. no. 212322). Antibacterial activity was assessed against Escherichia coli BW251 13 wild- type (Coli Genetic Stock Center #7636) and BW251 13 + pUC18-mcr-2. Additionally MIC90 assays were run on panels of COL-S (colistin-susceptible) clinical isolates collected from US sites between 2012 and 2016 for Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii (n=20/species) as well as a panel of COL-R (colistin-resistant) isolates (n=25, composed of a mixture of K. pneumoniae, P. aeruginosa and A. baumannii). Finally, E. coli ATCC 25922 WT and a panel of spontaneous mutants selected with Compound 14 and COL were assessed for changes in susceptibility to selecting drugs and a panel of compounds to assess cross-resistance.

MIC (Minimum Inhibitory Concentration) assays.

MIC assays were performed according to CLSI broth microdilution guidelines (M07-A9, M100-

S23) with the exception of using a 1 00 μί assay volume and preparing stock compounds at 50X final concentration. Briefly, stock solutions of all antibacterial agents were prepared fresh in appropriate solvents (i.e. DMSO, Dl water, ethanol, etc.). Stock concentrations were made at 50X the highest final assay concentration and serially diluted 2-fold, 12 times in a 96-well PCR plate (VWR cat. no. 83007- 374). Bacterial cell suspensions generated from MHA plate cultures were prepared in 0.85% saline and adjusted to -0.1 Οϋβοο nm. Next, cell suspensions were diluted 1 :200 in MHB (BD cat. no. 212322) to a concentration of ~5 x 10⁵ CFU (Colony-Forming Units)/mL. Ninety-eight microliters of each cell suspension in MHB were added to test wells in a 96-well assay plate (Costar cat. no. 3370). A Beckman Multimek 96 liquid handling robot was used to dispense 2 μί of each 50X stock compound into the plate containing 98 μΙ_ of each strain in MHB (2% final solvent concentration). Plates were shaken then incubated at 37°C overnight (16-20 h). MIC values were read visually at 100% growth inhibition for all compounds.

Part 2. Results

Table 13. Broth microdilution MICs ^g/mL) for Compounds 14 and 54a, PMB, and COL against Mcr-2- expressing E. coli

Tables 14A-14D. MICso and MIC90 analysis of Compounds 14 (CMP 14) and 54a (CMP 54a), PMB, COL, TIG (tigecycline), and MERO (meropenem) against COL-S E. coli (Table 14A), K. pneumoniae (Table 14B), P. aeruginosa (Table 14C), and A. baumannii (Table 14D) clinical isolate panels

Table 14A (COL-S E. coli)

MIC (Mg/mL)

# Strain CMP 14 CMP 54a PMB COL TIG MERO

3 MMX 8415 2 1 0.5 0.25 0.25 0.015

4 MMX 8416 2 2 0.5 0.5 0.25 0.015

5 MMX 8419 2 2 0.5 0.25 0.5 0.03

6 MMX 8421 2 1 0.5 0.5 0.25 0.015

7 MMX 8433 2 2 0.5 0.5 0.5 0.03

8 MMX 8880 2 1 0.5 0.25 0.25 0.015

9 MMX 8883 2 1 0.5 0.25 0.5 0.015 0 MMX 8885 2 1 0.5 0.5 0.25 0.015 1 MMX 8889 2 2 0.5 0.5 0.125 0.015 2 MMX 8890 2 1 0.5 0.25 0.25 0.015 3 MMX 8891 2 1 0.5 0.25 0.25 0.015 4 MMX 8915 2 1 0.5 0.25 0.125 0.015 5 MMX 8917 2 1 1 0.25 0.25 0.015 6 MMX 8920 2 1 0.5 0.5 0.25 0.015 7 MMX 8921 2 1 0.5 0.25 0.25 0.015 8 MMX 8923 2 1 0.25 0.5 0.25 0.015 9 MMX 8926 2 1 0.5 0.25 0.25 0.015 0 MMX 9013 2 1 0.5 0.5 0.25 0.015

MICso 2 1 0.5 0.25 0.25 0.015

MIC₉₀ 2 2 0.5 0.5 0.5 0.03 range 2-2 1-2 0.25-1 0.25-0.5 0.125-0.5 0.015-0.03

Table 14B (COL-S K. pneumoniae)

MIC (Mg/mL)

# Strain CMP 14 CMP 54a PMB COL TIG MERO 2 MMX 6379 4 2 1 0.25 1 0.03 3 MMX 6380 2 2 0.5 0.25 1 0.03 4 MMX 6381 2 1 0.5 0.25 1 0.03 5 MMX 6384 2 2 0.5 0.25 2 0.03 6 MMX 6386 2 1 0.5 0.5 1 8 7 MMX 8708 4 2 0.5 0.25 2 0.03 8 MMX 9030 2 1 0.5 0.25 1 0.06 9 MMX 61 17 4 2 0.5 0.25 1 0.06 0 MMX 6383 2 1 0.5 0.25 1 0.06

MICso 2 2 0.5 0.25 1 0.03

MIC₉₀ 4 2 0.5 0.5 2 0.5 range 2-8 1-2 0.5-1 0.25-0.5 1-8 0.03-8

Table 14C (COL-S P. aeruginosa)

MIC (Mg/mL)

# Strain CMP 14 CMP 54a PMB COL TIG MERO 0 MMX 8848 2 1 1 4 >8 1

MICso 2 1 1 1 >8 0.5

MIC₉₀ 4 2 1 2 >8 >8 range 2-8 1-2 0.5-1 0.5-4 >8->8 0.125->8

Table 14D (COL-S A. baumannii)

Table 14E. MICso and MIC90 analysis of Compounds 14 (CMP 14) and 54a (CMP 54a), PMB, COL, TIG, MERO, CAZ (ceftazidime), AMIK (amikacin), and LEVO (levofloxacin) against COL-R mixed-species clinical isolate panel

MIC (Mg/mL)

CMP CMP MER AMI

Species Strain PMB COL TIG CAZ LEVO

14 54a 0 K

K.

MMX

pneumoni 16 16 64 128 1 32 >256 16 32

6269

ae

K.

MMX

pneumoni 4 2 1 16 0.5 0.03 0.25 1 0.06

6382

ae

K.

MMX

pneumoni 4 4 64 256 2 64 >256 64 64

881 1

ae

P.

MMX

aeruginos 2 1 16 16 >8 1 4 4 0.5

7874

a

P.

MMX

aeruginos 8 8 256 1024 >8 8 64 32 >256

6977

a

A. MMX

2 1 16 256 4 128 >256 16 32 baumannii 8537

A. MMX

2 1 2 8 4 64 256 4 16 baumannii 8644

A. MMX

32 >32 64 256 4 128 256 8 16 baumannii 8990

A. MMX

2 1 8 128 4 64 >256 2 128 baumannii 6971

A. MMX >102 >25

4 2 256 2 16 32 8 baumannii 6973 4 6

A. MMX >25

2 1 8 32 4 32 128 8 baumannii 6975 6

A. MMX >25

4 2 32 512 4 64 >256 8 baumannii 8381 6

A. MMX >102 >25

8 8 256 4 8 128 16 baumannii 8383 4 6 MIC (Mg/mL)

CMP CMP MER AMI

# Species Strain PMB COL TIG CAZ LEVO

14 54a 0 K

A MMX >102 >25

23 4 2 256 2 16 64 8 baumannii 8384 4 6

A. MMX >25

24 4 1 8 64 2 32 128 8 baumannii 8386 6

A. MMX

25 2 2 2 8 2 32 128 4 16 baumannii 6338

A. MMX

26 2 ND^* ND 128 ND 128 ND ND ND baumannii 8537

MICso 4 2 32 128 2 32 >256 32 32

>102 >25

MIC₉₀ 16 16 256 8 128 >256 256

4 6

1- 8- 1-

0.5- 0.03- 0.25- 0.06- range 2-32 1->32 >102 >102 >25

>8 256 >256 >256 4 4 6

^* ND: not determined

Table 15. Summary of MIC90 analysis of Compounds 14 (CMP 14) and 54a (CMP 54a), PMB, COL, TIG, MERO, CAZ (ceftazidime), AMIK (amikacin), and LEVO (levofloxacin) against COL-S species and COL-R mixed-species clinical isolate panels

MIC₉₀ (range) ( g/mL)

CMP CMP

Species PMB COL TIG MERO CAZ AM IK LEVO

14 54a

>1024 128 >256 >256 256

COL-R 16 (2- 16 (1 - 256 (1 - 8 (0.5-

(8- (0.03- (0.25- (n=25) 32) >32) >1024) >8) (1 - (0.06- >1024) 256) >256) >256) >256)

Table 16. Summary of MIC90 analysis of Compound 84, Compound 90, Compound 91 , Compound 1 1 0, and COL, TIG and MERO against COL-S species and COL-R mixed-species clinical isolate panels

Example 113. Time-kill analysis of Compound 14 and colistin (COL) against E. coli ATCC 25922

Time-kill assays were performed according to CLSI guidelines (Methods for Determining Bactericidal Activity of Antimicrobial Agents; Approved Guideline. CLSI document M26-A (ISBN 1 -56238- 384-1 )). An overnight Mueller-Hinton agar (MHA) culture of E. coli ATCC 25922 was resuspended in 0.85% NaCI to -0.1 ODeoo and added to 10 mL Mueller-Hinton broth (MHB) medium to achieve a mid-10⁵ CFU/mL starting inoculum. Compound 14 and COL were then added to the inoculated MHB to produce concentrations equivalent to 0, 1 , 2, 4, and 16X the corresponding broth microdilution MIC value for each strain (Compound 14 - 2 μg/mL and COL - 0.5 μg/mL). Time-kills were performed in baffled, ventilated 125 mL Erlenmeyer flasks (TriForest, cat. no. FBC0125S) incubated 8ί 37 in a shaking incubator. Aliquots were removed from each flask at 0, 1 , 3, 6, 9, and 24 hours and serially diluted 1 :10 in 0.85% NaCI. Fifty microliters of each serial dilution were spread onto MHA plates and incubated for 24 hours at 37°C (assay limit of detection: 20 CFU/mL). Colonies were counted and CFU/mL values were calculated and plotted using GraphPad Prism 6 (GraphPad Software, Inc.) (FIGS. 3A and 3B). Fungistatic activity was defined by CFU reductions <3-logs from the starting inoculum and fungicidal activity was defined by CFU reductions≥3-logs (denoted by dashed lines in FIGS. 3A and 3B).

Similar assays were also performed using K. pneumoniae ATCC 43816, P. aeruginosa ATCC

27853, and A. baumannii ATCC 17978 and the results are shown in FIGS. 3C-3H.

Example 114. Spontaneous mutation frequency determination and cross-resistance analysis.

The spontaneous mutation frequency of Compound 14 and COL were determined for E. coli ATCC 25922. Three independent colonies of E. coli ATCC 25922 were picked from an overnight MHA plate and restreaked on fresh MHA plates and grown overnight. The following day a loop was used to scrape cells to generate an Ο βοο -0.80 suspension in 0.85% NaCI. One-hundred microliters (-6-8x10⁸ CFU) of each culture was then spread with sterile glass beads on plates (90 mm x 90 mm) containing 40 ml_ of MHA prepared with either 1 X or 2X the corresponding concentration of Compound 14 or COL needed to completely inhibit growth. Plates were incubated at 37°C for 48 hours and then putative mutant colonies were picked with a loop and streaked onto MHA plates containing the same

concentration of Compound 14 or COL that they were isolated from. Spontaneous mutation frequencies were then calculated by dividing the number of colonies that successfully regrew after 24 hours by the starting inoculum plated on each plate (Table 1 7). None of the 14 Compound 14-selected mutants and 2 of the 47 COL-selected spontaneous E. coli ATCC 25922 spontaneous mutants demonstrated broth microdilution MIC shifts 2-fold for the respective selecting compound (data not shown). The remaining 45 COL-selected mutants were evaluated in an MIC assay to determine the fold-shift changes vs. COL as well as cross-resistance to PMB, Compound 14, and MERO (Table 18). Table 17. Selection of Compound 14 and COL spontaneous mutants in E. coli ATCC 25922 on MHA plates containing 1 X or 2X the respective agar inhibition concentrations

All Compound 14 colonies selected grew upon restreaking on 1 X MIC agar media, however, when tested in broth microdilution MIC, failed to demonstrate≥2-fold reductions in susceptibility. ²AII but 2 of the COL-selected colonies at ^"I X or 2X MIC demonstrated≥2-fold reductions in susceptibility upon broth microdilution MIC testing.

Table 18. E. coli ATCC 25922 COL-R spontaneous mutants cross-resistance MIC panel (n=45 mutant strains)

Example 115. Membrane Permeability

The outer membrane of Gram-negative bacteria is composed of negatively charged LPS which constitutes a potent permeability barrier against the entry of hydrophobic compounds into cells. The ability of Compound 14 to damage the outer membrane was investigated using 1 -N-phenylnaphthylamine (NPN), an uncharged lipophilic dye. NPN fluoresces weakly in aqueous environments, but its

fluorescence greatly increases in hydrophobic environments such as the cell membrane. Therefore, the analysis of changes in fluorescence intensity of NPN in compound treated bacteria constitutes a sensitive and convenient method to evaluate changes in outer membrane permeability. Cell death was evaluated in parallel by use of SYTOX® Green, a green-fluorescent nuclear and chromosome counterstain that is impermeable to live cells.

Log-phase bacteria of two colistin-susceptible (E. coli ATCC25922 and K. pneumoniae 43816) and three colistin-resistant strains (K. pneumoniae MMX6949, P. aeruginosa MMX6977 and A. baumannii MMX8537) were used in the studies. Bacteria were treated with Compound 14, Compound 54a, Compound 59, or colistin at concentrations ranging from 0.1 to 5 μΜ in RPMI for 90 min. Compound 14, Compound 54a, and Compound 59 more potently induced membrane permeability for all bacteria when compared to COL (FIG. 4A). Bacterial cell death against COL^s susceptible strains was similar for Compound 14 and COL, whereas Compound 54a and Compound 59 killed all strains tested at lower concentrations (FIG. 4B). Treatment of COL^R strains with Compound 14 and Compound 54a resulted in significant cell death. These data demonstrate, Compound 14 and its analogs potently disrupt the outer membrane resulting in cell death in both COL^s and COL^R strains.

Damaging the outer membrane of Gram-negative bacteria has several important consequences:

(1 ) it can lead to direct killing of bacteria as shown by MIC assays and in the experiments described here,

(2) it sensitizes bacteria to killing by natural antimicrobials or in combination with other antibiotics, and (3) it results in increased complement fixation (data not shown) and CDC (complement dependent cytotoxicity).

Example 116. Antibacterial Activity of Compounds 14 against MDR CRE clinical isolates

Compound 14 was further evaluated against a panel of seven characterized multidrug resistant (MDR) carbapenem-resistant Enterobacteriaceae (CRE) clinical isolates, some of which possessed the mcr- 1 gene (Table 1 9). Similarly to the MDR isolates in the COL^R MIC90 panel, Compound 14 had consistent activity against these MDR CRE strains, including those possessing mcr- 1. Compound 14 had MICs of 2-4 μg/mL, except against a mucoid-producing strain (K. pneumoniae 30660 (Kp 30660)) where the MIC was 8 μg/mL. These data show the promising activity of Compound 14 against even the most problematic clinical isolates (such as the ceftazidime-avibactam resistant Klebsiella strain Kp 47772) independent of the potential added benefits from immune system engagement. Table 19

MOR fCs E co// 47554 Kf>¹ 4876S Kp 47768 K{ 47772 Kp 47640 Kp 30660* Kp 30684

ae

» .ν· ;^; ίϊ -Ϊ 'ίϊ, <¾¾»■^■,,,, Sfa

i.i¹. i^;f."'. i!,

■Λ¾. *iss Μκ<«.

<wf83. iSuft s .AS.

! W.

AO!SpKIS, iCfrttrtOS, vVT 0¾pC iCrsoi 3(> W OmsK¾6 ν'ί^'Γ Offif-KSe

MiC (Mg m!)

4 4

Meropenem 64 84 32 4 64 s 8

Cettaziiiitns-avibeetam >£4 1 4 266 >64 0.5 ·;

CeitaskSitnfc Hi4 >64 58 812 >S4 54 S4

Cefepitae >W >16 H28 s 16 18 S

2 >32 »258 8 32 32 32

^■ 64 NT-¹ NT 64 S4 6

Coiisiin 2 1 <2 <2 ■- 8 <C.5

Ttgeeye!me >4 1 NT NT 1 i

' Kp - KMmz!ls prsimo m; ⁵ mucoid producer; ·^■ OMP - Outer Mem&rane Protein; ⁴ Not Tes ed

Example 117. Compound Recognition by Rha Abs and Rha-Coordinated Binding to Bacteria

Rha-specific Abs (from vaccinated rabbits or natural Abs purified from human serum) were affinity purified and binding of Compound 14 to purified anti-Rha antibodies was measured using Surface Plasmon Resonance (BiaCore 3000). Binding was measured by flowing Compound 14 (analyte) over a CM5 chip coated with immobilized antibody (ligand). Equilibrium dissociation constants (KDs) of 87.3 nM and 66.8 nM were measured for complexes of Compound 14 with rabbit and human antibody, respectively. For Compound 59, which possesses two Rha molecules, KDs were 2.4 nM and 6.0 nM, respectively (data not shown).

The ability of Rha Ab to engage compound bound to bacteria was determined. ELISA plates were coated with 50 μΙ_ of a log-phase culture of E. coli ATCC 25922 (Ο βοο = 0.5)/well and dried.

Coated plates were then incubated with Compound 14, Compound 54a, and Compound 59 or compounds that do no have Rha molecules: Compound X and Compound Y (structrures shown further below) at indicated concentrations. Binding was then probed with purified human anti-Rha Ab at a fixed concentration. Bound antibodies were then detected using either an anti-human IgG secondary antibody (FIG. 5A) or an anti-human IgM secondary antibody (FIG. 5B). All three Rha-containing compounds (Compound 14, Compound 54a, and Compound 59) mediated binding of purified human Rha-Ab to E. coli. Binding was dose-dependent and both IgG (FIG. 5A) and IgM (FIG. 5B) isotypes were detected. To further demonstrate Rha-specific binding, competition assays using ELISA were performed with soluble L-rhamnose. As before, E. coli coated plates were incubated with various concentrations of Compound 14 and human anti-Rha Ab was added in the absence or presence (0.01 -1 mM) of L-rhamnose. Binding of Rha ab was detected using an anti-human IgG secondary antibody. L-rhamnose spike-in resulted in dose-dependent inhibition of anti-Rha Ab binding, confirming specificity for this carbohydrate (FIG. 5C). Furthermore, binding of purified rabbit or human anti-Rha Abs to six bacterial strains immobilized on ELISA plates was evaluated using Compound 14, Compound 54a, and Compound 59, colistin, and compounds that do no have Rha molecules: Compound X and Compound Y, as described above. ELISA plates were coated with 50 μί of a log-phase culture of the indicated bacterial strains (Ο βοο = 0.5)/ well and dried. Compounds were added at the indicated concentrations and probed with purified rabbit anti- Rha antibodies or human anti-Rha antibodies at a fixed concentration. Bound antibodies were detected using an anti-human IgG secondary antibody. Only Rha-containing compounds (Compound 14, Compound 54a, and Compound 59) mediated binding of purified rabbit (FIG. 6) or human anti-Rha abs (FIG. 7) to bacteria. Binding intensity was strain and dose-dependent. Binding of Rha antibodies to live bacteria was confirmed by flow cytometry (data not shown).

Collectively, these data demonstrate that L-rhamnose is effective at recruiting rabbit or human Rha antibodies to a wide variety of bacterial strains.

Compound X

Compound Y

Example 118. Rha-lnduced Enhancement of Bacterial Cytotoxicity in Whole Blood

In addition to possessing intrinsic antibiotic activity, the compounds may opsonize bacteria and recruit immune effectors to the site of infection. Using whole blood killing assays, immune-mediated enhancement of bactericidal activity has been demonstrated. 4x 10⁵ CFU/mL of E. coli 25922 were incubated in the presence or absence of Compound 14, Compound X, or colistin (COL) at sub-inhibitory concentrations (0.02 μΜ or about 0.03 μg/mL) in RPMI medium (FIGS. 8A and 8D) or human blood (FIGS. 8B and 8E). After 90 minutes of incubation, bacteria were harvested and colony forming units

(CFUs) were counted. The drug concentrations used were too low to inhibit bacterial growth in the absence of human blood (FIG. 8A, drug vs. RPMI alone). To correct for the intrinsic activity of human blood, the antibacterial activity of blood from donor 1 was measured - it reduced the CFUs/mL from 520k to 72k (86% reduction). Compound X or COL also improved antibacterial activity of blood. Compound 14 however, enhanced bacterial killing by more than tenfold (p<0.0001 , ANOVA with Tukey's adjustment), clearly demonstrating the Rha-mediated engagement of immune components present in blood (FIG. 8B, whole blood). This reduced the percentage of surviving bacteria from approximately 13% when treated with whole blood alone (or whole blood plus COL) to less than 1 % (FIG. 8C). The percentage survival was calculated relative to RPMI control for each compound to account for differences in non-Rha-induced antimicrobial activity. Results shown represent average with dots for individual data points and was analyzed by one-way ANOVA (^****P<0.0001 ). Donor 1 had anti-Rha Ab titers of 12,800 for IgG and 200 for IgM.

Using blood from another independent donor, killing enhancement with Compound 14 was retested, and compared to enhancement with Compound 54a and Compound 59. Killing enhancement observed with Compound 14 (donor 2, FIGS. 8D-8F) was similar to levels observed with donor 1 .

Compound 59 and Compound 54a were superior, enhancing bacterial killing by 4- and 1 0-fold, respectively, compared with Compound 14. It is important to note that anti-Rha IgG titers were lower in this particular donor (lgG=800, lgM=200) demonstrating that immune mediated killing enhancement is retained across a wide range of titers. Compound 54a and Compound 59 have consistently shown improved immune mediated activity as compared to Compound 14, demonstrating that the Rha-mediated activity of small molecule compounds can be optimized.

Of note, when mouse blood (spiked with rAb) was tested for whole blood killing using identical assay conditions, antibacterial activity was not observed in the presence or absence of compound (data not shown) highlighting key differences between mouse and human blood. One potentially important difference between human and mouse blood is white blood cell composition - human white blood cells contain -60% neutrophils, while much lower percentages are found in mouse blood. This is important when interpreting in vivo efficacy studies performed in mice as the immune-mediated component of bacteria killing may be underestimated in this species.

The human whole blood data presented above clearly demonstrate that the compounds benefit from immune-mediated enhancement of bactericidal activity. Importantly, multiple effector mechanisms potentially contribute to killing, including CDC, ADCC, and potentiation of the activity of antimicrobial peptides. The relative importance of respective mechanisms may vary in patients depending on individual predispositions. However, in cases where the pathogen displays resistance to antibiotic treatment, clinical outcomes may strongly depend on the successful interaction of these host immune mechanisms with the bacterium.

Example 119. Compound-Induced Enhancement of Complement-Dependent Cytotoxicity (CDC)

The complement system is one of the first lines of defense against invading pathogens, and plays an essential role in both innate and adaptive immunity. The complement cascade can be activated through distinct pathways; the classical pathway (CP), the lectin pathway (LP) or the alternative pathway (AP). The classical pathway is initiated by antibodies binding to the surface of the invading pathogen, while the alternative pathway acts through spontaneous hydrolysis of the protein C3. Whether initiated by either pathway, direct killing of pathogens by complement operates through the formation of the membrane attack complex (MAC). This activity can be measured in normal human serum (NHS) but is abolished in heat inactivated serum (HIS).

To demonstrate compound-mediated enhancement of complement activity, CDC assays were developed. Log phase E. Coli ATCC 25922 were generated by overnight culture and regrowth until OD₆oo=0.4 (1 e8 CFU/ml). 1 x 10⁶ CFU/mL of E. coli 25922 were incubated with sub-inhibitory

concentrations of 0.02 μΜ (-0.03 μg/mL) of Compond 14, Compound 54a or Compound 59, Compound X, or COL in RMPI alone (data not shown) or RPMI supplemented with 10% normal human serum (NHS) (FIGS. 9A and 9B) or 10% heat-inactivated (HIS) (FIGS. 9C and 9D). Plates were incubated at 37 °C for 90 minutes and surviving bacteria were determined by CFU enumeration (Rha titer lgG=6400 lgM=800). The low drug concentrations used did not inhibit bacterial growth in the absence of serum (RPMI alone, data not shown). Addition of NHS alone (FIGS. 9A and 9B) had little impact on bacterial growth compared to bacteria grown in RPMI (1 x 1 0⁶ to 4x 10⁶). Notably, while Compound X or COL displayed no antibacterial activity in NHS, Compound 14 enhanced bacterial killing by 22%, Compound 54a by 82%, and Compound 59 by 57% (FIG. 9B). This activity was not seen when the experiment was run with heat inactivated serum (HIS) (FIGS. 9Cand 9D), demonstrating that the effect was mediated by complement. These data derived from ex vivo killing assays suggest that the compounds with Rha attached recruit and activate both complement and cell-mediated immune effector arms of the immune system. Initial investigation of cell-mediated effects demonstrated the compounds increased neutrophil activity (data not shown).

Example 120. Mouse Models of Infection

The in vivo activity of Compound 14 has been evaluated in two key efficacy models, mouse septicemia (sepsis, bacteremia) and a more stringent neutropenic thigh infection model. Plans are to further evaluate Compound 14 in these models with other G- pathogens (K. pneumoniae, P. aeruginosa, A. baumanii, and Col^R pathogens) and in a lung infection model. The septicemia model is useful for compound screening and can provide two readouts, survival or kidney CFUs. Animals are infected IP and treated one hour later by the IP route. The thigh infection model, requires penetration of drug into muscle. The readout is thigh muscle CFUs. Animals are inoculated directly into the thigh and drug is dosed IP. It generally takes a higher dose of drug to demonstrate efficacy in the thigh versus the septicemia model. The success of Compound 14 in the immunocompromised thigh infection model supports its use in a non- immune-competent population.

Mice lack natural Rha Abs but they can be acquired by immunization or by passive transfer. Rha linked to ovalbumin provided a vaccine (OVA-R2) to immunize mice to raise anti-Rha antibodies.

However, Ab titers of mice immunized in this way were quite variable. To overcome this and obtain consistent titers, passive transfer of the antibodies was achieved by IV injection of affinity-purified antibodies isolated from the serum of OVA-R2 vaccinated New Zealand white rabbits (rAbs). Importantly, rAbs have been shown to bind murine Fc receptors as well as murine complement and, therefore, are expected to mediate an effect although the response may be less robust compared to a homogeneous system (i.e., a clinical setting).

Example 121. Immune-Competent Mouse Septicemia Model

Rha-mediated activity could be evaluated in vivo using an established, acute disseminated E. coli mouse septicemia model. Both intrinsic antibacterial activity of Compound 14 and additional immune- dependent efficacy relying on the presence of rAbs were investigated. Mice (n=12) infected with E. coli 25922 were given a single dose IP of Comppound 14 + rAb (FIG. 10,■), Compound 14 only (FIG. 10, o), or COL only.

FIG. 10 shows a dose ranging study with Compound 14 (MIC 2 μg/mL against the challenge strain) in a mouse E. coli septicemia model where kidney CFUs at 16 hours were enumerated. Improved efficacy was seen at 3 and 5 mpk in the presence of rAb with an additional ~1 -log reduction in burdens (P<0.05) versus Compound 14 in the absence of antibody or colistin (MIC 0.5 μg/mL) at 1 mpk. Plasma exposure of Compound 14 in this study was estimated at 2-3 μg^■hr/mL. Importantly, the bacterial burden in most mice in the "+rAb" groups was at the limit of detection (LOD) suggesting the actual effect may be significantly greater than it appears in the figure. In a similar study (data not shown), Compound X (no Rha attached) did not show rAb-enhanced efficacy while Compound 14 showed a dose-dependent reduction in CFUs which was improved in the presence of rAb. Example 122. Immune-Suppressed Mouse Thigh Infection Model

FIG. 1 1 shows a dose ranging study with Compound 14 (MIC 2 μg/mL) in a neutropenic mouse E. coli thigh infection model where bacterial burden in thigh muscle tissue at 24 hours was determined. Mice (n=6) infected with E. coli AT CC 25922 were administered Compound 14 either once (qd), twice (bid) or three times (tid) in one day either by IV (10 or 15 mg/kg/day), IP (20, 30, 60 or 90 mg/kg/day), or SC (20, 30, 60 or 90 mg/kg/day) routes. COL was administered IV qd (1 mg/kg/day) or IP qd (3 mg/kg/day) and gentamicin was administered SC bid (20 mg/kg/day). Only mice receiving Compound 14 were administered 1 00 μΙ of rabbit rAb 24h prior to E. coli challenge. Efficacy was demonstrated in all Compound 14 treated groups (+rAb), as well as in COL (MIC 0.5 μg/mL) and gentamicin (positive control) treated mice. E. coli thigh CFUs were significantly lower (p <0.01 ) in Compound 14, COL, and gentamicin treated mice. Compound 14 administered either IV, IP or SC bid or tid showed the greatest reduction in CFUs/thigh versus qd dosing. Higher variability seen in the Compound 14 SC treated mice was perhaps due to variable distribution to thigh tissues after SC dosing especially during the short duration of the study (24h endpoint). Compound 14 demonstrated robust efficacy at estimated plasma exposures (13.4 μg^■hr/mL) that are well below the NOAEL.

In a similar study activity of Compound 14 against K. pneumoniae in an immune-competent thigh model was also examined. In that study, an approximate 1 .5 log reduction in thigh burden was achieved when the animals were given a single IV dose of Compound 14 at 5 mg/kg (data not shown). Example 123. Antibody Titration Study

To investigate the minimum concentration of rAb necessary to engage the Rha of Compound 14 and produce an effect the concentration of rAb was varied in our standard E. coli Septicemia Model. Concentrations of rAb from (1000) to (32,000) in 4-fold increments (upper titrations not graphed for clarity; FIG. 12) were tested with Compound 14 fixed at 3 mg/kg (IP, single dose) and kidney burden elaborated at 16 hours. Mice (n=6) survived in all but the vehicle control group with 4 deaths.

As highlighted in FIG. 12, even at the lowest rAb concentration tested the Rha is fully engaged in this experiment and the maximum effect is achieved. However, it is worth noting that the majority of animals in the -i-rAb group are at the studies limit of detection, therefore it is possible that some differences in efficacy could be teased out if Compound 14 was tested at a lower concentration. Additional studies will be conducted to further clarify the low end of rAb concentration required for an effect in a murine system. It is encouraging that in our current test system potent efficacy is seen at modest Rha Ab titers.

Example 124. Drug Metabolism and Pharmacokinetics (DMPK) and Drug Stability

The plasma concentration-time profile and resulting pharmacokinetics of Compound 14 was determined upon dosing in mice (IV and IP; efficacy route), rats (IV and subcutaneous (SC); toxicology route), and cyno monkeys (IV and SC) (Table 20). It should be noted that the pharmacokinetics of Compound 14 (IP 10 mg/kg ± rAbs) was comparable with or without co-administered rAbs. The plasma half-life was approximately 1 .2 h, 0.5 and 0.6 h, in the mouse, rat and monkey, respectively. Plasma clearance in the respective animal species tested (mouse, rat, and monkey) appears to be low (<10% of corresponding liver blood flow). As plasma protein binding of Compound 14 was high (>99% bound) in all species including human, this could likely account for the overall lower clearance observed.

Table 20

AUCV* AUC_inf CL v„ V, F

Dose

Species Route (hr) (jig/inL)

(mg/kg) (μο-hr mL) (iiS-hi mi l (hr) (mL/miii/kg) (iuL/kg) (mL/kg) < )

Mouse IV bolus 3 NA 27.2 15.5 15.5 1.19 3.56 466 388 NA

IP 3 0.50 1.S0 2.09 2.30 NA NA NA NA 15%

IP 10 0.50 4.75 4.99 5.49 NA NA \ NA 1 1%

IP (+ rAbs) 10 1.00 2.74 3.78 6.72 NA NA NA NA 13%

SC 3 0.33 0.965 I 50 1.73 NA N A . NA 11%

Rat IV bolus 10 ^"MA 34.3 20.3 20.4 0.52 8.17 268 368 NA

SC 10 2.67 1.84 9.65 15.3 NA N A NA NA 75%

Monkey FV bohis 1 1.00 4.06 2.54 2.83 0.61 6.12 258 308

IV 1-hr inf 3 1.00 27.0 37.5 37.7 0.57 1.34 83.8 66.1 A

TV I-Iir inf 10 1.00 49.0 126 127 1.2 1.32 166 136

SC 10 1.17 25.1 77.7 88.8 MA NA. NA NA 70%

Bioavailability from SC dosing appeared to be high at about 70-75% in the species tested although IP dosing only generated a bioavailability of about 13% (repeated to confirm result). In the rat, parent drug was not detected in the urine. More recently, the plasma concentration-time profile of Compound 14 was characterized following a toxicokinetics study of IV 1 -hr infusion from a monkey tolerability study at doses of 3, 10, and 20 mg/kg. The resulting exposures suggest dose proportionality from 3 to 10 mg/kg. An earlier 1 mg/kg IV bolus administration was also studied.

Moreover, in vitro metabolic stability experiments carried out with un-labeled Compound 14 incubations in whole blood and liver microsomes/hepatocytes indicate that the compound is stable. FIG. 13 shows the metabolic stability of Compound 14 in rat, monkey and human hepatocytes.

Furthermore, from in vivo studies, we have further profiled for metabolites employing

LC-QTof-MS mass defect filtering in select plasma/urine samples from the rat and have not detected any Ph1 or oxidative biotransformation products. We had also specifically looked for precursors of the parent compound that are available as synthetic standards, including those that involve cleavage of the linker, but have not found any to date. Therefore, the bifunctional compound appears to be stable in vivo. Thus far, there does not appear to be significant metabolism and it may be that we will need to characterize its metabolism using radio-labeled compound. We are also characterizing the concentrations of Compound 14 in various tissues. Example 125. Off-Target Activity

Compound 14 was evaluated in the CEREP SafetyScreen44 battery of in vitro

enzyme/receptor/ion channel assays at 10 μΜ. Six CNS targets were inhibited at 55-85% (a2A adrenergic, κ-opioid, 5-HTI B, 5-HT2a receptors and dopamine and norepinephrine transporters) and of lesser concern since Compound 14 is unlikely to cross the blood brain barrier due to its multiple charge state and size. Screening against a broader panel is planned. Binding affinity (ICso) to the rat potassium A-type channel (KA), a neuronal target, was 330 nM; COL had a ICso of 9610 nM. The binding assay results were followed up in functional human voltage-gated potassium channels Kv1 .1 , Kv1 .2 and Kv1 .6 using an electrophysiological platform (lonWorks Quattro). Results from this functional assay suggest that antagonism of the channels occurred at much higher concentrations than observed in the binding assay with ICso values estimated to be >100 μΜ (FIG. 14). Minimal CYP450 inhibition (1 A2, 2B6, 2C8, 2C9, 2C19, 2D6, 3A4) was observed when tested at a range of Compound 14 concentrations with ICso values determined to be >30 μΜ.

Example 126. Toxicology

Compound 14 in 5-Day Rat Toxicology Study. COL induces renal toxicity in humans at clinically relevant doses. Compound 14 was evaluated for nephrotoxicity using a rat model predictive of COL-induced renal injury in humans. Compound 14 was administered SC for 5 days to rats at 25 mg/kg/day BID, which generates plasma exposures (-24 μg^■hr/mL), 12-fold higher than exposures required for efficacy in the mouse septicemia model. COL was similarly administered at 25 mg/kg/day, a dose that generates therapeutic plasma levels in humans. Urine was collected daily and monitored for albumin levels, a biomarker for renal proximal tubule injury. In contrast to COL, Compound 14 did not elevate urinary albumin. Lack of renal toxicity with Compound 14 was confirmed microscopically in kidneys collected on Day 6 (data not shown). Compound 14-treated rats demonstrated no adverse clinical pathology, hematology, or histopathology (major organs). Acute histamine-like effects were seen (similar to COL) but were rat specific; there were no adverse observations in singly-dosed cynos.

Histamine-like effects are not expected in humans, as none are seen with COL. Since these effects are likely Cmax-related, slow infusion of drug may minimize any potential histamine-related responses. Example 127. Compound 14 Safety Margins

Based on the more stringent thigh infection model that utilized an E. coli strain with an MIC of 2 μg/mL, results from the 5-day rat toxicology and single dose cyno tolerability studies, as well as exposures from mouse, rat, and cyno PK studies, the following margins have been calculated (Table 21 ). Based on projected efficacious plasma AUCs (10 mg/kg, bid, IP) and exposures as outlined below, there is a≥2.9-fold margin established regarding nephrotoxicity (likely to be wider) and a >19-fold margin regarding neuromuscular effects. Now that the effect in cynos has been correlated with Cmax, various dosing protocols will be explored (tid, 2h infusion or continuous infusion) to allow better toxicological assessment using higher exposures.

Table 21

Example 128. Synergistic Activity of of Compounds 14 and 54a

Part 1. Methods

Checkerboard synergy assays.

The synergistic activity of Compound 14 in combination with Gram-positive agents (AZITHRO

(azithromycin) and RIF (rifampicin)) and Gram-negative agents ((TIG (tigecycline), MERO (meropenem), LEVO (levofloxacin), AMIK (amikacin), and CAZ/AVB (ceftazidime/avibactam)) was assessed for the seven COL-R clinical isolates possessing the highest MIC values for Compound 14 (e.g.,≥8 μς/ιηί). Checkerboard synergy assays were performed as previously described (see, e.g., Eliopoulos and Moellering, "Antibiotics in Laboratory Medicine," 4th Edition (The Williams & Wilkins Co., Baltimore, Maryland, 1996), and Moody, "Clincal Microbiology Procedures Handbook," 2nd Edition, Volume 2 (Washington: ASM Press, 2004)). In master stock plates, the stocks of two compounds (Compound A, which is Compound 14 in this Example; and Compound B, which is a known antibacterial agent) for synergistic evaluation were prepared. A liquid stock of Compound A was prepared fresh in Dl water to a concentration 50X the desired final synergy assay concentration. Into wells A1 through H1 (vertical, Y- axis column on left edge of plate) of a 96-well PCR plate (VWR cat. no. 83007-374) Compound A was aliquoted in 100 μΙ_ volumes. To all other wells on the plate, with the exception of A12 through H12, 50 μΙ_ of Dl water was added. A multichannel pipette was then used to transfer 50 μΙ_ from column A1 -H1 to column A2-H2. Following mixing, tips were discarded and the process was repeated across the plate to create a series of 1 1 , serial two-fold dilutions spanning columns 1 through 1 1 .

A stock of Compound B was prepared in a similar fashion in the appropriate solvent (DMSO, Dl water, ethanol, etc.) at 50X the desired final synergy assay concentration. In a separate 96-well PCR plate, 100 μΙ_ aliquots of Compound B were added to wells in the top row of the plate spanning A1 through A12. In all other wells, excluding the bottom row (H1 -H12), 50 μΙ_ of the corresponding

Compound B solvent were added. In a similar fashion to Compound A, Compound B was serially diluted, two-fold down the plate using a multichannel pipette, resulting in 7 rows comprised of 12 wells with 50 μΙ_ diluted Compound B. In some cases a second dilution plate, setup in the same fashion as the first, was needed to capture synergy present at drug concentrations less than the range on the first plate.

Checkerboard synergy assays were performed using the same assay setup as the standard MIC assays previously described previously herein (i.e. inoculum, media, volume, 96-well assay plates, incubation time, incubation temperature, 100% inhibition assay endpoint, etc.). However, rather than a single addition of 2 μΙ_ of 50X stock compound solution to the assay plate containing MHB pre-inoculated with bacterial cells using the Multimek 96 liquid handling robot, two sequential additions were made (one from the Compound A 50X stock plate and one from the Compound B 50X stock plate). The resulting checkerboard plates contained a 7 x 1 1 -well grid (A1 -G1 x A1 -A1 1 ) of synergy wells containing varying proportions of Compounds A and B, an 1 1 -point dilution series of Compound A alone (wells H1 -H1 1 , assay concentration range 1 X to 1 /1024X), a 7-point dilution series of Compound B alone (wells A12- G12, assay concentration range 1 X to 1 /64X) and a positive growth control well containing no drug (well H12) (FIG. 1 ).

FIC Compound A = MIC of Compound A in combination/MIC of Compound A alone

FIC Compound B = MIC of Compound B in combination/MIC of compound B alone

Part 2. Results

Table 22. Summary of checkerboard synergy analysis for Compound 14 in combination with Gram-positive and Gram-negative standard of care agents for COL-R isolates possessing the highest

MIC values for Compound 14

MIC (Mg/mL)

K_i pneumonae.

6⁹⁵¹

Kⁱ pneumonae.

Compound 14 alone 16 1 6 MMX ⁶⁹⁵⁶ 16 8 8 4 32

Compound 14 combe ) 4 ί 4 4 4 4 2

AZITHRO alone 256 25 56 Kⁱ pneumonae.128 512 128 1024 1024

AZITHRO combo 32 3 2 8 MMX ⁶⁹⁶¹ 32 64 1024 1

FICI 0.38 0.. 38 0.31 0.56 1 2 0.06

Compound 14 alone 32 3 2 8 Kⁱ pneumonae. 8 4 8 64

Compound 14 combe ) 4 > 2 2 MMX ⁶²⁶⁹ 4 2 2

RIF alone 16 6 4 32 32 32 2 2

Pⁱ aerugnosa.

RIF combo 0.125 i 4 8 32 0.125 0.125

6⁹⁷⁷

FICI 0.13 0. 13 0.38 0.5 2 0.31 0.09

Compound 14 alone 32 1 6 16 8 8 A bⁱⁱaumann. 4 64

Compound 14 combe ) 8 i 8 8 4 2 MMX ⁸³⁸³ 4

TIG alone 8 ί > 2 1 16 4 4

A bⁱⁱaumann.

TIG combo 2 1 0.5 1 2 2 0.125

8⁹⁹⁰

FICI 0.5 0.^" 75 0.75 2 0.63 1 0.09

Compound 14 alone 16 1 6 8 8 4 4 64

Compound 14 combe ) 8 1 6 4 4 4 2 2

MERO alone 32 1∑ Ϊ8 32 32 8 8 64

MERO combo 8 Y >8 4 4 4 1 1

FICI 0.75 : » 0.63 0.63 1 0.63 0.05

Compound 14 alone 16 1 6 16 8 8 4 64

Compound 14 combe ) 4 ί 4 0.125 4 2 2

LEVO alone 256 25 56 64 32 1024 8 16

LEVO combo 32 6 4 32 16 256 2 0.25

FICI 0.38 0 5 0.75 0.52 0.75 0.75 0.05

Compound 14 alone 16 1 6 8 8 8 8 64 MIC (Mg/mL)

Compound 14 comb o Kⁱ pneumonae 0..031 4 0.031 4 0.5 4 16

AMIK alone 64 MMX ⁶⁹⁵¹ 32 32 16 64 1024 4

AMIK combo 32 16 16 8 32 0.016 1

Kⁱ pneumonae.

FICI 0.5 0.75 0.5 1 0.56 0.5 0.5

6⁹⁵⁶

Compound 14 alons 3 16 16 8 16 8 8 64

Compound 14 comb o 8 8 1 4 4 2 8

Kⁱ pneumonae.

CAZ/AVB alone 8 8 16 8 8 64 64

6⁹⁶¹

CAZ/AVB combo 2 1 8 4 1 4 16

FICI 0.75 0.63 0.63 0.75 0.63 0.31 0.38

Kⁱ pneumonae.

6²⁶⁹

Example 129. Synthesis of INT-42 (fe -butyl (2S)-2-aminooctanoate)

Pⁱ aerugnosa.

6⁹⁷⁷

A suspension of (S)-2-aminooctano c acid (3.2 g, 20 mmol) in tert-butyl acetate A bⁱⁱaumann. (30 mL) was cooled in an ice-water bath and added with perchloric acid 70 % (4.4 g). After the mixture w MMX ⁸³⁸³as stirred at 0 °C to room temperature for 2.5 days, it was concentrated by rotary evaporation. The remaining was diluted with MeOH (5 mL) and poured into a beaker containing ice (50 g) and NaOH pellet (2.3 g) A bⁱⁱ.aumann A. fter all the NaOH was consumed, the pH was adjusted to -7.5 with solid Na2C03. The suspension was MMX ⁸⁹⁹⁰ extracted by DCM (1 00 mL x 5). The combined organic layers were dried over Na2S04, concentrated by rotary evaporation and further dried under high vacuum. Yield 1 .93 g, 44.8 %. Ion found by LCMS: [M- tBu + H]+ = 160.

Example 130. Synthesis of INT-43

Step a. Synthesis of methyl (15S)-1-[(6-deoxy-alpha-L-mannopyranosyl)oxy]-15-hexyl-11-(2-{[(2S)- 1-methoxy-1-oxooctan-2-yl]amino}-2-oxoethyl)-7,10,13-trioxo-3-oxa-6,11 ,14-triazahexadecan-16- oate

A mixture of methyl (2S)-2-aminooctanoate (490 mg, 2.8 mmol) and INT-39 (600 mg, 1 .29 mmol) was dissolved in anhydrous DMF (2 mL) and DIPEA (702 mg, 5.4 mmol). The solution was cooled in an ice-water bath followed by dropwise addition of a solution of HATU (1 .06 g, 2.8 mmol) in DMF (3 mL) via syringe pump at a rate of 1 .5 ml/hr. After the addition, the reaction was directly purified using RPLC (100 g, 5 to 100 % acetonitrile and water). Yield 528.7 mg, 52.8 %. Ion found by LCMS: [M + H]⁺ = 777.

Step b. Synthesis of (15S)-11-(2-{[(1S)-1-carboxyheptyl]amino}-2-oxoethyl)-1-[(6-deoxy-alpha-L- mannopyranosyl)oxy]-15-hexyl-7,10,13-trioxo-3-oxa-6,11 ,14-triazahexadecan-16-oic acid

A solution of step-a product (528.7 mg, 0.681 mmol) in a 1 :1 mixture of MeOH:THF (6 mL) was cooled in an ice-water bath. It was added with a solution of LiOH (40 mg, 1 .67 mmol) in water (3 mL). The mixture was stirred for 3 hours, then neutralized by 4N HCI solution in dioxane. After partial concentration, the reaction was directly purified by RPLC (100 g, 0 to 80 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 487.4 mg, 95.6 %. Ion found by LCMS: [M + H]⁺ = 749.

706-Linker A (more polar, by HPLC RT) and 707-Linker B (less polar, by HPLC RT)

Step a. Coupling of L-nor-Leu Methyl Ester and Removal of Boc Group

HATU (3.1 g, 8.1 mmol) in DMF (5 mL) was added, dropwise, to a solution of racemic-transl - (ferf-butoxycarbonyl)pyrrolidine-3,4-dicarboxylic acid (1 g, 3.9 mmol), L-Norleu-OMe-hydrochloride (1 .5 g, 8.1 mmol), and triethylamine (4.0 g, 39.5 mmol) in DMF (10 mL) over a period of 20 minutes. The mixture was stirred for an additional 20 minutes then applied directly to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 1 0% to 95% acetonitrile and water using 0.1 % TFA modifier. The two diastereomers were separated and pooled and lyophilized separately into the more polar diastereomer (a) and the less polar diastereomer (b): LC/MS [M-Boc+H]+ = 414.2 for both Boc-protected intermediates. Each diastereomer was stirred in a 1 /1 mixture of DCM/TFA (8 mL) for 20 minutes then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 0% to 75% acetonitrile and water using no modifier to afford the TFA salt after lyophilization as a hydroscopic white solid. Yield: 88%, 2 steps. LC/MS [M+H]+ = 414.2.

Note: The stereochemistry of the trans-pyrrolidine dicarboxylate diastereomers was arbitrarily assigned based on HPLC retention time.

Step b. Cbz Protection and Hydrolysis of Methyl Esters

Cbz-NHS ester (227 mg, 0.91 mmol) was added to a stirring mixture of the product from Step a

(more polar diastereomer) (400 mg, 0.76 mmol) and triethylamine in DMF (10 mL). The mixture was stirred for 4 hours and then applied to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 5% to 95% acetonitrile and using 0.1 % THFA modifier. The solvent was removed by rotovap and the di-ester was hydrolyzed stirring in a 1 /1 /2 mixture of methanol/THF/DI water containing LiOH (0.72 g, 30 mmol) for 30 minutes. The mixture was adjusted to pH 5 with acetic acid then concentrated and applied to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 10% to 95% acetonitrile and water using 0.1 % TFA modifier. The pure fractions were lyophilized to afford INT-44a as a white solid. Yield: 78%, 2 steps. LC/MS [M+H]+ = 520.2.

Example 132. Synthesis of INT-44

HATU (644mg, 1 .69 mmol), in DMF (2 mL) was added, dropwise over 10 minutes, to a stirring mixture of INT-18 (2.65 g, 1 .69 mmol), INT-44a (400 mg, 0.77 mmol), and triethylamine (466 mg, 4.62 mmol) in DMF (6 mL) and the reaction was stirred for an additional 30 minutes. Acetic acid ( 2 mL) was added, followed by 5% Pd/C (400 mg) and the mixture was stirred under an atmosphere of hydrogen for 12 hours. The mixture was filtered through celite and applied to reversed phase liquid chromatography (RPLC) using an Isco Combiflash liquid chromatograph eluted with 35 % to 95% acetonitrile and water using 0.1 % TFA modifier. Yield: 35%. LC/MS [M+3H/3]+ =1 092.8.

Example 133. Preparation of INT-45

Tri-tert-butyl {[(2S,5R,8S,1 1 S,14S,17S,22S)-22-({(8S,1 1 S,14R)-8-amino-14-{2-[(tert- butoxycarbonyl)amino]ethyl}-1 1 -[(1 R)-1 -hydroxyethyl]-2,2-dimethyl-4,9,12,15-tetraoxo-3-oxa-5,10,13- triazapentadecan-15-yl}amino)-5-benzyl-17-[(1 R)-1 -hydroxyethyl]-8-(2-methylpropyl)-3, 6,9,12,15,18,23- heptaoxo-1 ,4,7,10,13,16,19-heptaazacyclotricosane-2,1 1 ,14-triyl]tri(ethane-2,1 -diyl)}triscarbamate was prepared by an analogous procedure to the preparation of INT-18 except D-Dab was utilized instead of L- Dab. Positive mass ion was found as (M - Boc + 2H⁺)/2: 732.5), tr = 1 .89 minutes with 5 min gradient (40-95%) method.

Example 134. Preparation of INT-46. Trans-3-({4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanamido}methyl)cyclopropane-1 ,2- dicarboxylic acid (pair of diastereomers)

(pair of diastereomers) Step a: Synthesis of dimethyl trans-3-(hydroxymethyl)cyclopropane-1 ,2-dicarboxylate

To a solution of aldehyde (see Preparation of INT-26, Step c, 4.496 g, 24.15 mmol) in THF (177 ml_) in a 1000 ml_ round bottom flask was added glacial AcOH (30 ml_, THF:AcOH = 177:30) and NaBh CN subsequently at 0 °C. The reaction was warmed up to room temperature for 15 min. Water (177 ml_) was added and pH was adjusted to 7 with saturated NaHCCb solution. The aqueous layer was extracted with ether (3 x 100 mL) and the combined organic layers were dried over Na2S04. Solvents were removed. Silica gel column chromatography (0.5-6% MeOH/DCM, UV@214nm) to give the alcohol 2.85 g (63%). Ή NMR (300 MHz, Chloroform-d) δ 3.99 (dd, J = 1 1 .9, 5.4 Hz, 1 H), 3.84 (dd, J = 12.5, 7.4 Hz, 1 H), 3.75 (s, 3H), 3.73 (s, 3H), 2.38 (d, J = 7.4 Hz, 2H), 2.14 (qd, J = 7.4, 5.4 Hz, 1 H).

Step b: Synthesis of dimethyl trans-3-(bromomethyl)cyclopropane-1 ,2-dicarboxylate

To a stirred solution of alcohol (2.85 g, 15.1 mmol) in CH2CI2 (130 mL) at room temperature were added PPh₃ (6.02 g, 22.7 mmol) and CBr₄ (7.70 g, 22.8 mmol). After being stirred for 1 hr, the mixture was concentrated under reduced pressure and the residue was purified by column chromatography (10-15% Et20/Hexanes, UV@214nm) on silica gel to give bromide as a colorless oil (3.58 g, 94%). ¹³C NMR (100 MHz, Chloroform-d) δ 170.8, 1 69.7, 52.5, 52.4, 30.4, 29.8, 28.5, 28.3

Step c: Synthesis of dimethyl trans-3-(azidomethyl)cyclopropane-1 ,2-dicarboxylate

To a stirred solution of bromide (3.58 g, 14.3 mmol) in DMF (0.1 5 M) at room temperature were added TBAI (1 1 .7 g, 31 .1 mmol) and NaN₃ (4.68 g, 71 .3 mmol). After stirring at 80 °C for 2 hrs., the mixture was cooled to room temperature and then treated with H2O (two volumes). The mixture was extracted with hexane (3x). Combined organic layer was dried over Na2S04 and concentrated under reduced pressure to give the azide as a colorless oil (2.96 g, 97%), which was pure enough and was used for the next reaction without purification: Rf = 0.2 (Et2O/hexane=10:90); ¹ H NMR (300 MHz, Chloroform-d) δ 3.76 (s, 3H), 3.74 (s, 3H), 3.68-3.48 (m, 2H),2.43 (dd, J = 9.2, 4.8 Hz, 1 H), 2.30 (t, J = 5.4 Hz, 1 H), 2.21 -2.08 (m, 1 H).

Step d: Synthesis of dimethyl trans-3-{[(tert-butoxycarbonyl)amino]methyl}cyclopropane-1 ,2- dicarboxylate

To a stirred solution of the azide (2.96 g, 13.9 mmol) in THF (20 mL) and MeOH (40 mL) at room temperature were added B0C2O (15.3 g, 69.4 mmol) and Pd(OH)2 (2.43 g, 3.47 mmol). After stirring under a hydrogen balloon for 3 hrs, the mixture was filtered through a pad of Celite and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (ether/hexane = 20:80 - 60:40) to give N-Boc amine as a colorless oil (3.31 g, 83%): Rf = 0.33

(ether/hexane = 30:70); Ή NMR (300 MHz, Chloroform-d) δ 4.71 (bs, 1 H), 3.74 (s, 3H), 3.72 (s, 3H), 3.60 - 3.56 (m, 1 H), 3.39 - 3.27 (m, 1 H), 2.34 (dd, J = 9.2, 4.8 Hz, 1 H), 2.25 (dd, J = 5.4 Hz, 1 H), 2.13 (d, J = 7.5 Hz, 1 H), 1 .46 (s, 9H). Step e: Synthesis of dimethyl ester of INT-46. Dimethyl trans-3-({4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanamido}methyl)cyclopropane-1 ,2- dicarboxylate (a pair of diastereomers)

The Boc amine (0.180 g, 0.627 mmol) was treated with 4NHCI in dioxane (4 mL) for 1 hour followed by removing excess HCI and dioxane. The amine HCI salt and INT-2 (0.330 g, 0.940 mmol) were dissolved in DMF 10 mL followed by adding DIPEA (0.16 mL, 0.94 mmol). HATU (0.357 g, 0.940 mmol) was added into the reaction mixture at once. After 1 hr, DMF was removed and the residue was loaded into reversed phase prep-HPLC (0-1 5%ACN/Water with 0.1 %TFA) to give 0.238 g (73%). Positive mass ion was found as (M + H⁺): 521 .2, tr = 2.53 minutes with 8 min (5-95%) method.

Step f : Synthesis of INT-46. Trans-3-({4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanamido}methyl)cyclopropane-1 ,2- dicarboxylic acid (a pair of diastereomers)

Dimethyl ester of INT-46 (0.238 g, 0.457 mmol) was mixed with LiOH (0.0237 g, 0.960 mmol) in 6 mL of THF/Water (1 :1 ). Stirred for less than 1 hrs. 1 N HCI to acidify. Removed THF and extracted with EtOAC to give INT-46 (0.220 g, 97%). Positive/negative mass ions were found as (M + H⁺): 493.2, (M - H⁺): 491 .2, tr = 0.52 minutes with 5 min (5-40%) method.

INT-47 was prepared using a similar procedure as that for INT-16 except D-Dab is used in place of L-Dab. Positive mass ions were found as (M - Boc + 2H⁺)/2: 632.4 (M + H⁺): 1364.1 , tr = 1 .41 minutes with 5 min (40-95%) method. Example 136. Preparation of INT-48

INT-47 (0.904 g, 0.663 mmol), Z-L-2-amino-octanoic acid (0.214 g, 0.729 mmol) and DIPEA (0.17 ml_, 0.99 mmol) were dissolved in 10 mL of DMF. HATU (0.378 g, 0.995 mmol) was added as DMF solution (5 mL) via syringe pump. Stirred till the reaction completed. Added 5% Pd/C (0.423 g, 0.1 99 mmol) and stirred under hydrogen balloon for 3 hours. Removed DMF and purified with reversed phase prep-HPLC to give INT-48 (0.806 g, 81 %). Positive mass ions were found as (M - Boc + 2H⁺)/2: 703.0, tr = 4.72 minutes with 8 min (5-95%) method.

Example 137. Preparation of INT-49. Trans-3-{(1 E)-3-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-3-oxoprop-1-en-1-yl}cyclopropane-1 ,2-dicarboxylic acid and trans-3-{3-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-3- oxopropyl}cyclopropane-1 ,2-dicarboxylic acid (pair of trans-cyclopropyl diastereomers)

Step a: Synthesis of di-tert-butyl trans-3-[(1 E)-3-methoxy-3-oxoprop-1-en-1-yl]cyclopropane-1 ,2- dicarboxylate

The sulfonium salt (see INT-26 step a for procedure, 6.000 g, 24.88 mmol)) was added into a flask with 50 mL of THF at 0 °C. t-BuOK (1 M THF, 24.9 mL, 24.9 mmol) was added and stirred for 20 min, then at -78 °C, di-tert-butyl fumarate (4.982 g, 20.73 mmol) and 20 mL of THF were added respectively. The reaction mixture was stirred at -78 °C for 5 hrs and then let warm-up to room temperature. After the reaction was completed the reaction was quenched with brine (60 mL). The organic layer was separated and the aq. layer was extracted with ether twice. Dried and 0-15% Hex/EA silica column purification to give the desired product 0.983 g (15%). ¹³C NMR (75 MHz, Chloroform-d) δ 169.49, 167.78, 166.09, 143.1 1 , 123.53, 82.03, 81 .82, 77.47, 77.04, 76.62, 51 .45, 30.29, 29.91 , 29.57, 28.02, 27.99.

Step b: Synthesis of (2E)-3-[trans-2,3-bis(tert-butoxycarbonyl)cyclopropyl]prop-2-enoic acid

The product of Step a (0.212 g, 0.650 mmol) was mixed with LiOH (0.0400 g, 1 .63 mmol) in 6 mL of THF/Water (1 :1 ). The reaction mixture was stirred for less than three days. 1 N HCI was added to acidify the reaction mixture. Removed THF and extracted with EtOAC. Purified by reversed phase prep- HPLC to give the desired acid 0.170 g (83%). HPLC showed tr = 7.2 min for the product. Step c: Synthesis of di-tert-butyl ester of INT-49 and INT-49a. Di-tert-butyl trans-3-{(1 E)-3-[(2-{2- [(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-3-oxoprop-1-en-1-yl}cyclopropane- 1 ,2-dicarboxylate (mixture of diastereomers)

The acid (0.140 g, 0.448 mmol) and INT-1 (0.135 g, 0.538 mmol) were dissolved in 10 mL of DMF followed by adding DIPEA (0.26 mL, 0.67 mmol). HATU (0.256 g, 0.672 mmol) was added into the reaction solution at once. After 1 hr, DMF was removed and the residue was loaded into reversed phase prep-HPLC (0~15%ACN/Water with 0.1 %TFA) to give the di-tert-butyl ester of INT-49, 0.207 g (85%). Positive mass ion was found as (M + H⁺): 546.2, tr = 4.30 minutes with 8 min (5-95%) method.

Step d: Synthesis of INT-49. Trans-3-{(1 E)-3-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-3-oxoprop-1-en-1-yl}cyclopropane-1 ,2-dicarboxylic acid

The di-tert-butyl ester (0.200 g, 0.367 mmol) was mixed with 6 mL of TFA and stirred for less than 1 hrs. Removed TFA and purified reversed phase prep-HPLC to give INT-49 (mixture) (0.130 g, 83%). Positive/negative mass ions were found as (M + H⁺): 434.2, (M - H⁺): 432.2, tr = 2.1 1 minutes with 5 min (5-40%) method. The product was obtained as a mixture of trans-cyclopropyl dicarboxylate

diastereomers. Example 138. Preparation of INT-50

The synthesis of INT-50 was analogous to that for INT-20 except D-Dab was used in place of L- Dab in the synthesis. Positive mass ion was found as (M - 2Boc + 2H+)/2: 803.3, tr = 2.44 minutes with 5 min (40-95%) method.

Example 139. Preparation of INT-51 and INT-52

Step a: Hydrogenation of di-tert-butyl trans-3-[(1 E)-3-methoxy-3-oxoprop-1-en-1-yl]cyclopropane- 1 ,2-dicarboxylate

The olefin (see INT-49, Step a for a procedure, 2.00 g, 6.13 mmol) in 40 mL of THF was treated with 5% Pd/C. The mixture was degassed by bubbling argon through the solution and then it was stirred under a H2 atmosphere (balloon pressure), the reaction mixture was stirred for 2h. Celite filtration and solvent removal gave the clean desired product in 1 00% yield. ¹ H NMR (300 MHz, Chloroform-d) δ 3.69 (s, 3H), 2.30 - 1 .95 (m, 2H), 2.15 (ddd, J = 9.5, 4.7, 0.7 Hz, 1 H), 2.09 - 1 .95 (m, 2H), 1 .94 - 1 .78 (m, 1 H), 1 .77 - 1 .63 (m, 1 H), 1 .48 (s, 9H), 1 .46 (s, 9H).

Step b: Synthesis of 3-[trans-2,3-bis(tert-butoxycarbonyl)cyclopropyl]propanoic acid Mixed the di-tert-butyl ester (2.012 g, 6.13 mmol) with LiOH (0.182 g, 7.35 mmol) in 32 mL of THF/Water (1 :1 mL). The mixture was stirred for about three days. 1 N HCI was added to acidify the reaction mixture. Removed THF and extracted with EtOAC to give the desired acid 1 .926 g (100%). ¹ H NMR (300 MHz, Chloroform-d) δ 2.43 (t, J = 7.4 Hz, 2H), 2.17 (dd, J = 9.4, 4.7 Hz, 1 H), 2.14 - 1 .95 (m, 2H), 1 .96 - 1 .84 (m, 1 H), 1 .83 - 1 .68 (m, 1 H), 1 .48 (s, 9H), 1 .46 (s, 9H).

Step c: Synthesis of di-tert-butyl trans-3-{3-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-3-oxopropyl}cyclopropane-1 ,2-dicarboxylate (a pair of diastereomers)

The acid (1 .926 g, 6.13 mmol) and INT-1 (1 .693 g, 6.74 mmol) were dissolved in 10 mL of DMF followed by adding DIPEA (1 .60 mL, 9.1 9 mmol). HATU (3.494 g, 9.19 mmol) was added into the reaction solution at once. After 1 hr, DMF was removed and the residue was loaded into reversed phase prep- HPLC (0~15%ACN/Water with 0.1 %TFA) to give 2.002 g (60%). Positive mass ions were found as (M + H⁺): 548.2 and (M - CeHnOs + H⁺): 400.2, tr = 4.46 minutes with 8 min (5-95%) method.

Step d: Synthesis of trans-3-{3-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]- 3-oxopropyl}cyclopropane-1 ,2-dicarboxylic acid (a pair of diastereomers)

Mixed the di-tert-butyl ester (2.000 g, 3.65 mmol) with TFA (30 mL). Stirred for less than 1 hrs. Removed TFA followed by adding water and lyophilization to give the diacid (100%). Negative mass ion was found as (M - H⁺): 434.2, tr = 2.20 minutes with 5 min (5-40%) method.

Step e: Synthesis of INT-51 and INT-52

The diacid (1 .590 g, 3.652 mmol) and methyl (2S)-2-aminooctanoate HCI salt (1 .608 g, 7.668 mmol) were dissolved in 10 mL of DMF/DCM (1 :4) followed by adding NaHCC (1 .227 g, 14.61 mmol), HOAt (1 .242 g, 9.129 mmol) and EDCI (1 .750 g, 9.129 mmol). After 3hrs, the reaction mixture was diluted with EtOAc (100 mL) and washed with 1 N HCI, saturated NaHCCb and brine. Solvents were removed and the residue was loaded into reversed phase prep-HPLC (0-1 5%ACN/Water with 0.1 %TFA) to give INT- 51 as Peak-1 (polar) and INT-52 as Peak-2 (less polar). Positive/negative mass ions were found as (M + H⁺): 747.2 and (M - CeHnOs + H⁺): 600.4, (M - H⁺): 745.0, tr = 4.84 minutes with 8 min (5-95%) method for INT-51 and (M + H⁺): 747.2 and (M - CeHnOs + H⁺): 600.4, (M - H⁺): 745.0, tr = 5.37 minutes with 8 min (5-95%) method for INT-52. The stereochemistry of the trans-cyclopropane was arbitrarily assigned.

Example 140. Preparation of INT-53 N~1 ~,N~31 ~-bis(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)-4,7,10,13,19,22,25,28-octaoxa-16-azahentriacontane-1 ,31- diamide and INT-54 1-[(6-deoxy-alpha-L-mannopyranosyl)oxy]-22-{21-[(6-deoxy-alpha-L- mannopyranosyl)oxy]-15-oxo-3,6,9, 12,19-pentaoxa-16-azahenicosan-1 -yl}-7,23-dioxo- 3,10,13,16,19-pentaoxa-6,22-diazahexacosan-26-oic acid

Step a: Synthesis of 16-[(benzyloxy)carbonyl]-4,7,10,13,19,22,25,28-octaoxa-16-azahentriacontane- 1 ,31-dioic acid

NH-(PEG4-COOH (1 .441 g, 2.806 mmol) was dissolved in 1 5 mL of THF and 10%aq. Na₂C0₃ (15 mL) and treated by CbzCI (0.62 mL, 4.2 mmol) by dropwise addition with ice-water bath cooling. After stirring overnight, the mixture was acidified with 1 N HCI and THF/water was removed followed by adding minimal amount of NMP/water. Prep-HPLC purification gave the desired Cbz protected diacid 1 .53 g, 84%. Positive/negative mass ions were found as (M + H⁺): 648.4, (M - H⁺): 646.2, tr = 3.92 minutes with 8 min (5-95%) method.

Step b: Synthesis of INT-53. N~1 ~,N~31 ~-bis(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)-4,7,10,13,19,22,25,28-octaoxa-16-azahentriacontane-1 ,31- diamide

The Cbz protected diacid (1 .53 g, 2.36 mmol), INT-1 (1 .247 g, 4.961 mmol), and DIPEA (1 .03 mL, 5.91 mmol) were mixed in DMF (20mL). HATU (2.245 g, 5.905 mmol) in DMF (13 mL) was pumped into the mixture in 4hrs. The reaction was stirred for another hour after the completion of HATU addition. DMF was removed and the residue was purified by prep-HPLC to give the Cbz protected INT-53, 2.142 g, 81 %. Positive mass ion was found as (M - 2C6H11 O5 - Bn + H⁺): 698.4, tr = 4.37 minutes with 8 min (5-95%) method. The Cbz protected INT-53 was dissolved in ethanol and treated with 5%Pd/C and hydrogen balloon for 2hrs. Celite filtration and solvent removal gave INT-53 1 .88 g, 100%. Positive mass ions were found as (M - 2C₆Hn 04 + 2H⁺)/2: 344.6, (M - CeH C + 2H⁺)/2: 417.6.

Step c. Synthesis of INT-54

INT-53 (1 .156 g, 1 .18 mmol), succinic acid monobenzyl ester (0.2937 g, 1 .41 1 mmol), and DIPEA (0.61 mL, 3.5 mmol) were mixed in DMF (1 0 mL). HATU (0.671 g, 1 .76 mmol) in DMF (6 mL) was pumped into the mixture in 4hrs. The reaction was stirred for another 1 hr after the completion of HATU addition. DMF was removed and the residue was purified by prep-HPLC to give 1 .05 g, 76%. The benzyl ester was dissolved in ethanol and treated with 5% Pd/C and hydrogen balloon for 2hrs. Celite filtration and solvent removal gave INT-54 which was used for next step reaction without purification. Positive mass ions were found as (M - 2C₆Hi i0₄ + 2H⁺)/2: 394.8, (M - C₆Hii0₄ + 2H⁺)/2: 467.8, (M + 2H⁺)/2: 540.8. Example 141. Preparation of INT-55 and INT-56

Step a. Synthesis of INT-55. Methyl (4S,12S)-4,12-dihexyl-3,6,10-trioxo-2-oxa-5,8,11- triazatridecan-13-oate

2,2'-{[(benzyloxy)carbonyl]azanediyl}diacetic acid (0.9994 g, 3.740 mmol), methyl (2S)-2- aminooctanoate HCI salt (1 .647 g, 7.854 mmol), EDCI (1 .792 g, 9.350 mmol), HOAt (2.273 g, 9.350 mmol) and NaHCCb (1 .257 g, 14.96 mmol) were mixed in 20 mL of DCM/DMF (4:1 ). The reaction mixture was stirred for 3hrs. EtOAc was added into the reaction mixture to dilute followed by washing with 1 NHCI, saturated NaHC03 and brine wash. After removal of solvents the residue was loaded into normal phase prep-HPLC, 0.5~7%MeOH/DCM to purify. 2.008 g of the desired product (Cbz protected INT-55) was obtained, 93%. Rf = 0.6 7% MeOH/DCM. ¹ H NMR (300 MHz, Chloroform-d) δ 8.25 (d, J = 7.6 Hz, 1 H), 7.42 (d, J = 7.9 Hz, 1 H), 7.39 - 7.22 (m, 5H), 5.08 - 5.20 (m, 2H), 4.65 - 4.52 (m, 1 H), 4.51 - 4.39 (m, 1 H), 4.00 (d, J = 5.5 Hz, 4H), 3.72 (s, 3H), 3.67 (s, 3H), 1 .58 - 1 .90 (m, 4H), 1 .40 - 1 .15 (m, 16H), 0.87 (t, J = 19.7 Hz, 6H, 2CH3). ¹³C NMR (75 MHz, CDC ) δ 172.99, 172.90, 169.35, 169.28, 155.85, 135.97, 128.44, 128.05, 127.66, 67.91 , 53.32, 53.20, 52.57, 52.55, 52.53, 52.27, 52.13, 32.10, 31 .68, 31 .54, 28.81 , 25.47, 25.25, 22.53, 14.02. The Cbz protected INT-55 was dissolved in ethanol and treated with 5%Pd/C and hydrogen balloon for 2hrs. Celite filtration and solvent removal gave INT-55 used for next step reaction without purification. Positive/negative mass ions were found as (M + H⁺): 444.4, (M - H⁺): 442.2, tr = 3.95 minutes with 8 min (5-95%) method.

Step b. Synthesis of INT-56

INT-54 (0.26 g, 0.228 mmol), INT-55 (0.127 g, 0.228 mmol), and DIPEA (0.079 mL, 0.46 mmol) were mixed in 5 mL of DMF. HATU was added via syringe pump (2 mL, 1 mL/hr). The reaction solution was stirred for another hour. DMF was removed and the residue was purified by prep-HPLC to give 0.224 g, 65%. Positive mass ions were found as (M - 2 CeH C + 2H⁺)/2: 607.6, (M - CeH C + 2H⁺)/2: 680.4, (M + 2H⁺)/2: 753.6, tr = 2.1 0 minutes with 5 min (40-95%) method. Example 142. Preparation of INT-57 (15S)-11-(2-{[(1S)-1-carboxyheptyl]amino}-2-oxoethyl)-1-[(6- deoxy-alpha-L-mannopyranosyl)thio]-15-hexyl-7,10,13-trioxo-3-oxa-6,11 ,14-triazahexadecan-16-oic acid

Per-acetated INT-9 (0.0730 g, 1 .48 mmol), INT-55 (0.0656 g, 0.148 mmol) and DIPEA (0.052 mL, 0.30 mmol) were mixed in 4 mL of DMF. HATU (0.0844 g, 0.222 mmol) was added via syringe pump (1 mL, 1 mL/hr). The reaction solution was stirred for another hour after completion of addition. DMF was removed and the residue was purified by prep-HPLC to give Per-acetated dimethyl ester of INT-57, 0.083 g, 61 %. Positive mass ion was found as (M - CeH C + H⁺): 647.0, tr = 3.46 minutes with 5 min (40-95%) method. Per-acetated dimethyl ester of INT-57 (0.083 g, 0.090 mmol) was dissolved in 6 mL of THF/water (1 :1 ) and treated with LiOH (0.01 1 g, 0.46 mmol). After acidification and ethyl acetate extraction, clean INT-57 was obtained in 100%. Positive/negative mass ions were found as (M + H⁺): 765.4, (M - H⁺): 763.4, tr = 4.10 minutes with 8 min (5-95%) method.

Example 143. Preparation of INT-58 and INT-59

Step a. Synthesis of INT-58

The Cbz protected diacid (see INT-53, Step a for procedure, 0.200 g, 0.309 mmol), INT-53 (0.6355 g, 0.6484 mmol), and DIPEA (0.22 ml_, 1 .2 mmol) were mixed in DMF (1 OmL). HATU (0.294 g, 0.772 mmol) in DMF (3 mL) was pumped into the mixture in 4hrs. The reaction was stirred for another hour after the completion of HATU addition. DMF was removed and the residue was purified by prep-

HPLC to give the Cbz protected INT-58, 0.660 g, 83%. Positive mass ions were found as (M - 4CeHn04 + 3H⁺)/3: 662.9, (M - 3C₆Hii0₄ + 3H⁺)/3: 71 1 .4, (M - 2C₆Hii0₄ + 3H⁺)/3: 760.3, (M - CeHnC + 3H⁺)/3: 809.0, tr = 3.28 minutes with 8 min (5-95%) method. The Cbz protected INT-58 was dissolved in ethanol and treated with 5%Pd/C and hydrogen balloon for 2hrs. Celite filtration and solvent removal gave INT-58 1 .88 g, 1 00%. Positive mass ions were found as (M - CeH C + 2H⁺)/2: 667.0, (M - 2C₆Hn04 + 3H⁺)/3: 716.0, (M - C₆Hii04 + 3H⁺)/3: 764.6, (M + 3H⁺)/3: 813.4, (M - CeHnC + 2H⁺)/2: 1 146.4, (M + 2H⁺)/2: 1219.2.

Step b. Synthesis of 4-(bis{2-[(1-methoxy-1-oxooctan-2-yl)amino]-2-oxoethyl}amino)-4- oxobutanoic acid

Succinic acid monobenzyl ester (0.568 g, 2.73 mmol), INT-55 (1 .10 g, 2.48 mmol) and DIPEA (1 .30 mL, 7.44 mmol) were mixed in 20 mL of DMF. HATU (1 .414 g, 3.72 mmol) was added via syringe pump (10 mL, 2mL/hr). The reaction solution was stirred for another hour after the completion of addition. DMF was removed and the residue was purified by prep-HPLC to give dimethyl benzyl triester, 1 .245 g, 79%. Positive mass ion was found as (M + H⁺): 634.4, tr = 2.85 minutes with 7 min (60-95%) method.

Dimethyl benzyl triester (1 .245 g, 1 .964 mmol) was dissolved in ethanol and treated with 5% Pd/C and H2 balloon for 2 hrs. Celite filtration gave the desired acid in 1 00%. Positive mass ion was found as (M + H⁺): 544.0, tr = 3.01 minutes with 5 min (40-95%) method. Step c. Synthesis of INT-59

The acid from Step b (0.0950 g, 0.175 mmol), INT-58 (0.387 g, 0.159 mmol) and DIPEA (0.083 mL, 0.47 mmol) were mixed in 4 mL of DMF. HATU (0.0901 g, 0.238 mmol) was added via syringe pump (2 mL, 1 mL/hr). The reaction solution was stirred for another hour after the completion of addition. DMF was removed and the residue was purified by prep-HPLC to give dimethyl ester of INT-59, 0.31 8 g, 66%. Positive mass ions were found as (M - 4C₆Hn04 + 3H⁺)/3: 793.4, (M - 3CeHn04 + 3H⁺)/3: 842.3, (M -

2C₆Hii0₄ + 3H⁺)/3: 890.3, (M - CeHnC + 3H⁺)/3: 939.6, (M + 3H⁺)/3: 987.9, tr = 4.18 minutes with 8 min (5-95%) method. Dimethyl ester of INT-59 (0.318 g, 0.107 mmol) was dissolved in 8 mL of THF/water (1 :1 ) and treated with LiOH (0.0054 g, 0.23 mmol). After acidification THF was removed and 3mL of NMP was added. Then most of Water was removed and the residue was load to pep-HPLC to purify. 0.289 g of INT-59 was obtained (92%). Positive mass ions were found as (M - 3C₆Hn04 + 4H⁺)/4: 624.8, (M

4C₆Hii0₄ + 3H⁺)/3: 784.0, (M - 3C₆Hi i0₄ + 3H⁺)/3: 832.8, (M - 2C₆Hii0₄ + 3H⁺)/3: 881 .8, (M - CeHnC + 3H⁺)/3: 929.8, (M + 3H⁺)/3: 979.8, tr = 3.80 minutes with 8 min (5-95%) method. Example 144. Preparation of INT-60. 2,2'-{[(2-{3-[(3,4,5-Trihydroxy-6-methyloxan-2- yl)oxy]propanamido}acetamido)acetyl]azanediyl}diacetic acid

Step a. Synthesis of 3-[(2,3,4-tri-0-acetyl-6-deoxyhexopyranosyl)oxy]propanoic acid

To a solution of the per-acetated bromo-L-rhamnopyranose (1 .600 g, 4.531 mmol) and benzyl 3- hydroxypropanoate (0.6280 g, 3.485 mmol) in 50 mL of dry CH2CI2 at room temperature were added silver triflate (1 .075 g, 4.182mmol) and 4A molecular sieves (12 g). After the mixture was stirred for 2 hrs. at room temperature. Two Grams of Celite was added into the reaction mixture followed by filtration. The filtrate was condensed and purified with 1 -5% MeOH/DCM to give the desired product 0.853 g, 54%. ¹ H NMR (300 MHz, CDCI3) δ 7.36, 7.35, 7.34, 7.32, 7.31 , 7.31 , 7.29, 7.29, 7.28, 5.25, 5.24, 5.22, 5.21 , 5.20, 5.20, 5.1 9, 5.14, 5.08, 5.06, 5.04, 5.03, 5.00, 4.73, 4.73, 4.02, 4.00, 3.99, 3.98, 3.97, 3.95, 3.92, 3.90, 3.89, 3.88, 3.87, 3.86, 3.85, 3.83, 3.73, 3.71 , 3.70, 3.69, 3.67, 3.66, 3.44, 2.67, 2.65, 2.63, 2.13, 2.12, 2.04, 2.02, 1 .98, 1 .96, 1 .30, 1 .20, 1 .18. ¹³C NMR (75 MHz, CDC ) δ 170.89, 170.01 , 169.95, 169.87, 135.68, 128.56, 128.24, 128.21 , 97.60, 77.57, 77.1 5, 76.72, 71 .00, 69.70, 69.06, 66.51 , 66.44, 63.37, 34.59, 20.84, 20.76, 20.67, 17.34. The benzyl ester was hydrogenated (5% Pd/C, H₂ balloon) to the free acid in quantitative yield.

Step b. Synthesis of N-{3-[(2,3,4-tri-0-acetyl-6-deoxyhexopyranosyl)oxy]propanoyl}glycylglycine The propanoic acid (from Step a, 0.6830 g, 1 .885 mmol), NH₂-Gly-Gly-OBn HCI salt (0.4877 g,

1 .885 mmol), EDCI (0.5420 g, 2.828 mmol), HOAt (0.3849 g, 2.828 mmol) and NaHCOs (0.6334 g, 7.540 mmol) were mixed in 1 0 mL of (DCM/DMF (4:1 ). The reaction solution was stirred overnight. The reaction mixture was diluted with EtOAc and washed with 1 NHCI, saturated NaHCCb and brine. Reversed phase prep-HPLC purification gave the desired product 1 .020 g, 95%. Positive mass ion was found as (M + H⁺): 567.0, tr = 4.23 minutes with 8 min (5-95%) method. The benzyl ester was hydrogenated (5% Pd/C, H₂ balloon) to the free acid in quantitative yield.

Step c. Synthesis of INT-60

The acid (from Step b, 0.8410 g, 1 .782 mmol), dimethyl 2,2'-azanediyldiacetate (0.3837 g, 1 .942 mmol) and DIPEA (0.46 mL, 2.6 mmol) were mixed in 10 mL of DMF. To the solution, was added HATU (1 .007 g, 2.648 mmol) in DMF (5 mL) with syringe pump at 1 mL/min. The reaction solution was stirred for another hour after completion of addition. Reverse phase prep-HPLC purification gave the desired product, dimethyl ester of INT-60, 1 .016 g, 93%. Positive mass ion was found (M + H⁺): 61 9.8, tr = 3.36 minutes with 8 min (5-95%) method. Dimethyl ester of INT-60 (1 .016 g, 1 .640 mmol) was dissolved in 20 ml_ of THF/water (1 :1 ) and treated with LiOH (0.3927 g, 16.40 mmol). After acidification THF was removed and 3ml_ of NMP was added. Then most of Water was removed and the residue was load to pep-HPLC to purify. 0.567 g of INT-60 was obtained (74%). Negative mass ions were found as (M - C₆Hi i 04 - H⁺): 302.0, (M - H⁺): 464.0, tr = 0.80 minutes with 5 min (5-60%) method.

Example 145. Preparation of INT-61. 2,2'-{[(2-{(R)-3-[(3,4,5-Trihydroxy-6-methyloxan-2- yl)oxy]butanamido}acetamido)acetyl]azanediyl}diacetic acid

Step a: Synthesis of (R)-3-[(2,3,4-tri-0-acetyl-6-deoxyhexopyranosyl)oxy]butanoic acid

To a solution of the per-acetated bromo-L-rhamnopyranose (1 .900 g, 5.380 mmol) and benzyl (R)-3-hydroxybutanoate (0.8708 g, 4.483 mmol) in 80 mL of dry CH2CI2 at room temperature were added silver trifiate (1 .256 g, 4.887 mmol) and 4A molecular sieves (20 g). After the mixture was stirred for 2 hrs. at room temperature. 2 gram of Celite was added into the reaction mixture followed by filtration. The filtrate was condensed and purified with 1 -5% MeOH/DCM to give the desired product 1 .018 g, 49%.¹ H NMR (300 MHz, CDCI3) 7.33 (s, 2H), 7.38 - 7.23 (m, 2H), 5.35 - 5.08 (m, 4H), 5.02 (t, J = 9.9 Hz, 1 H), 4.84 (d, J = 1 .8 Hz, 1 H), 4.34 - 4.17 (m, 1 H), 3.98 - 3.82 (m, 1 H), 2.65 (dd, J = 15.5, 8.0 Hz, 1 H), 2.47

(dd, J = 15.5, 4.9 Hz, 1 H), 2.14 - 1 .92 (m, 7H), 1 .17 (dd, J = 10.1 , 6.2 Hz, 5H). ¹³C NMR (75 MHz, CDCI3) δ 170.83, 170.12, 1 69.96, 1 69.86, 135.67, 128.53, 128.19, 94.73, 77.61 , 77.18, 76.76, 71 .03, 70.30, 69.48, 69.10, 66.62, 66.39, 42.00, 20.85, 20.66, 18.84, 17.26.The benzyl ester was hydrogenated (5% Pd/C, H2 balloon) to the free acid in quantitative yield.

Step b. Synthesis of N-{(R)-3-[(2,3,4-tri-0-acetyl-6-deoxyhexopyranosyl)oxy]butanoyl}glycylglycine

The (R)-3-butanioc acid (from Step a, 0.8120 g, 2.1 58 mmol), NH₂-Gly-Gly-OBn HCI salt (0.5582 g, 2.158 mmol), EDCI (0.6204 g, 3.236 mmol), HOAt (0.4405 g, 3.236 mmol) and NaHCOs (0.7250 g, 8.630 mmol) were mixed in 10 mL of (DCM/DMF (4:1 ). The reaction solution was stirred overnight. The reaction mixture was diluted with EtOAc and washed with 1 NHCI, saturated NaHCCb and brine. Reverse phase prep-HPLC purification gave the desired product 1 .224 g, 98%. Positive mass ion was found as (M + H⁺): 581 .2, tr = 4.38 minutes with 8 min (5-95%) method. The benzyl ester was hydrogenated (5% Pd/C, H2 balloon) to the free acid 1 .01 1 g, 98%. Positive/negative mass ions were found as (M + H⁺): 491 .2, (M - H⁺): 489.2, tr = 2.45 minutes with 5 min (5-95%) method. Step c. Synthesis of INT-61

The acid (from Step b, 1 .01 1 g, 2.061 mmol), dimethyl 2,2'-azanediyldiacetate (0.4481 g, 2.268 mmol) and DIPEA (0.54 mL, 3.1 mmol) were mixed in 10 mL of DMF. To the solution, was added HATU (1 .176 g, 3.092 mmol) in DMF (5 mL) with syringe pump at 1 mL/min. The reaction solution was stirred for another hour after completion of addition. Reverse phase Prep-HPLC purification gave the desired product, dimethyl ester of INT-61 , 1 .173 g, 90%. Positive mass ion was found (M + H⁺): 633.8, tr = 3.45 minutes with 8 min (5-95%) method. Dimethyl ester of INT-61 (1 .173 g, 1 .851 mmol) was dissolved in 20 mL of THF/water (1 :1 ) and treated with LiOH (0.4434 g, 18.51 mmol). After acidification THF was removed and 3mL of NMP was added. Then most of Water was removed and the residue was load to pep-HPLC to purify. 0.773 g of INT-61 was obtained (87%). Negative mass ions were found as (M - H⁺): 478.0, tr = 0.60 minutes with 5 min (5-60%) method.

Example 146. Preparation of INT-62

INT-62

INT-62 was prepared from INT-1 1 and Z-D-Ser-OH in an analogous manner as described in Example 14. Yield: 81 %. LC/MS ((M+2H)/2) = 525.6 (loss of 1 Boc group).

Step a. Synthesis of the Protected Octapeptide

A solution of HATU (1 .97 g, 5.18 mmol) in DMF (9 mL) was added via a syringe pump at a rate of 2.5 mL/hr into a solution of INT-1 1 (5.0 g, 4.71 mmol), Z-D-Ser-OH (1 .24 g mg, 5.18 mmol),

diisopropylethylamine (1 .82 g, 14.12 mmol) in DMF (35.5 mL). After the addition, the reaction mixture was stirred for an additional hour then purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 35 to100% methanol and water to yield the desired compound (6.05 g, 1 00%). Ion found by LC/MS [(M-2Boc+2H)/2]+ = 542.4. Step b. Synthesis of INT-62

To a solution of the product from Step a. in methanol (31 mL) was charged with 5% Pd/C (94.2 mg) and H2 gas balloon. The reaction was stirred at room temperature overnight. Next day, the reaction mixture was filtered through a pad of Celite ® and washed with methanol then concentrated. The residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 78% methanol and water to yield the title compound as a free base (2.61 g, 48%). Ion found by LC/MS [(M-2Boc+2Na)/2] = 497.6.

Example 147. Preparation of INT-63

INT-63 was prepared from INT-62 and Z-Thr-OH in an analogous manner as described i e 16 (Procedure A). Yield: 66%. LC/MS ((M+2HV2) = 525.8 (loss of 2 Boc groups).

Example 148. Preparation of INT-64

Step a. Synthesis of the Protected Dipeptide

The synthesis of the protected dipeptide was accomplished by a similar procedure using L- threonine methyl ester hydrochloride (1 .0 equiv.), Z-Dab(Boc)-OH dcha (1 .05 equiv.), HOBt (1 .5 equiv.), EDCI (1 .5 equiv.), and NaHCCb (2.0 equiv.) to yield the methyl ester. Hydrolysis of the methyl ester with LiOH (3.0 equiv.) in MeOH:THF:H₂0 (1 :1 :2) to yielded the desired compound (51 %). Ion found by LC/MS [M-H]+ = 452.2. Step b. Coupling of the Dipeptide to INT-62

A solution of HATU (218.34 mg, 0.57 mmol) in DMF (1 mL) was added via a syringe pump at a rate of 2.5 mL/hr into a solution of 836-INT-B (0.6 g, 0.52 mmol), 836-INT-E (260.41 g mg, 0.57 mmol), diisopropylethylamine (202.25 mg, 1 .57 mmol) in DMF (6 mL). After the addition, the reaction mixture was stirred for an additional hour then purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 35 to 100% methanol and water, using 0.1 % TFA as modifier to yield the title compound (577.0 mg, 66%). Ion found by LC/MS [(M-2Boc+2H)/2]+ = 694.5.

Step c. Synthesis of INT-64

A solution of the Step-b Product (51 1 .0 mg, 303.09 mmol) in MeOH (6 mL) was charged with 5% Pd/C (96.76 mg, 0.045 mmol) and H2 gas balloon. The reaction mixture was stirred at room temperature overnight. After the reaction was complete, it was filtered through a pad of Celite®, washed with methanol then concentrated. The residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 30 to 100% methanol and water to yield INT-64 as the free base (191 .0 mg, 43%). Ion found by LC/MS [(M-3Boc+3H)/3]+ = 385.4.

Example 149. Preparation of INT-65

INT-65 was prepared from INT-64 and Z-L-Norleu-OH in an analogous manner as described for INT-62. Yield: 67%. LC/MS [M+2H/2] 682.6 (loss of 2 Boc groups).

xample 150. Preparation of INT-66

derived from more polar intermediate

Note: The absolute configuration of the trans-pyrrolidne dicarboxamide is arbitrarily assigned but the product is derived from the more polar trans-cyclopentane-(bis-octanoate methyl ester)

Step a. Synthesis of dimethyl (3R,4R)-1-benzylpyrrolidine-3,4-dicarboxylate

To a solution of dimethyl fumarate (1 .44 g, 10 mmol) in acetonitrile (40 mL) was added N- (methyoxymethyl)-N-(trimethylsilylmethyl)benzylamine (2.37 g, 10 mmol) followed by lithium fluoride (410 mg, 15.8 mmol). After the mixture was stirred for 1 .5 days, it was concentrated by rotary evaporation. The residue was purified by RPLC (150 g, 0 to 35 % acetonitrile and water, using 0.1 % TFA as modifier). The collected was concentrated by rotary evaporation and further dried in high vacuum to afford the product as a yellow oil TFA salt. Yield 3.0 g, 76.7 %. Ion found by LCMS: [M + H]⁺ = 278.

Step b. Synthesis of (3R,4R)-1-benzylpyrrolidine-3,4-dicarboxylic acid

The step-a product (3 g, 7.67 mmol) was dissolved in 1 :1 mixture of MeOH/THF (16 mL). The solution was cooled in an ice-water bath and added with a solution of LiOH (384 mg, 15.3 mmol) in water (10 mL). After the reaction mixture was stirred overnight, it was added with an additional amount of LiOH (300 mg) in water (3 mL) and continued at ~ 5-10 °C for 6 hours. 4N HCI solution in dioxane (3 mL) was slowly added to adjust pH to ~ 6. The organic solvents was removed by rotary evaporation, and the aqueous solution was purified by RPLC (150 g, 0 to 50 % acetonitrile and water). Yield 1 .57 g, 82 %. Ion found by LCMS: [M + H]⁺ = 250. Step c. Synthesis of dimethyl (2S,2'S)-2,2'-{[(3R,4R)-1-benzylpyrrolidine-3,4- diyl]bis(carbonylazanediyl)}dioctanoate

A mixture of step-b product (498.6 mg, 2 mmol) and methyl (2S)-2-aminooctanoate HCI (922.7 mg, 4.4 mmol) was dissolved in anhydrous DMF (4 mL) and DIPEA (1 .69 g, 13 mmol). The resulting mixture was added with a solution of HATU (1 .67 g, 4.4 mmol) in DMF (5 mL) via syringe pump at a rate of 5 ml/hr. After the addition, the reaction was directly purified by RPLC (1 50 g, 10 to 70 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 540 mg of the desired product as more polar isomer, 48.2 %. Ion found by LCMS: [M + H]⁺ = 560.

Step d. Removal of the benzyl protection

The step-c more polar isomer (540 mg, 0.96 mmol) was dissolved in MeOH. The solution was added with Pd(OH)2 and stirred under hydrogen for 20 hours. Pd(OH)2 was filtered off, and the filtrate was concentrated to dryness. Yield 450 mg, 99.8 %. Ion found by LCMS: [M + H]+ = 470.

Step e. Synthesis of dimethyl (2S,2'S)-2,2'-{[(3/?,4/¾-1-{4-[(2-{2-[(6-deoxy-a-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoyl}pyrrolidine-3,4- diyl]bis(carbonylazanediyl)}dioctanoate

A mixture of the step-d product (450 mg, 0.958 mmol) and INT-2 (405.5 mg, 1 .15 mmol) was dissolved in anhydrous DMF (2 mL) and DIPEA (390 mg, 3 mmol). After the solution was cooled in an ice- water bath, it was drop-wise added with a solution of HATU (437 mg, 1 .15 mmol) in DMF (1 mL) at a rate of 2 ml/hr. After the addition, the reaction was stirred for 30 more minutes and purified by RPLC (150 g, 10 to 70 % acetonitrile and water). Yield 250 mg, 32.4 %. Ion found by LCMS: [M + H]⁺ = 803. Step f. Synthesis of INT-66

The step-e product (250 mg, 0.31 mmol) was dissolved in a 1 :1 mixture of MeOH:THF (4 mL). After the solution was cooled in an ice-water bath, it was added with a solution of LiOH (32 mg, 1 .34 mmol)) in water (2 mL) by three portions over 1 .5 hours and stirred for one more hour. It was then acidified by 4N HCI solution in dioxane (0.3 mL) and purified by RPLC (50 g, 5 to 55 % acetonitrile and water, using 0.1 % TFA as modifier). The yield of INT-66 was 171 mg, 68.1 %. Ion found LCMS: [M + H]⁺ = 775.

The dicarboxylic acid INT-26 (0.042 g, 0. 85 mmol) and INT-45 (0.266 g, 0.1 70 mmol) were dissolved in 5 mL of DMF followed by addition of DI PEA (0.037 ml_, 0.21 mmol). To this mixture was added HATU (0.0808 g, 0.213 mmol) in 1 .5 mL of DMF via syringe pump over 2 hours at room temperature. After completion of HATU addition, the reaction mixture was stirred for another hour at room temperature. Then the reaction mixture was concentrated under reduced pressure. The residue was purified by reversed phase prep-HPLC with ACN/water (0.1 %TFA) to give the per-Boc-Compound 61 as Peak-1 (polar) and per-Boc-Compound 62 as Peak-2 (less polar). The mass spectrum showed strong detectable positive charge signals at tr = 2.19 and 2.43 min with 7 min (60-95%) method [found positive ion (M - 5Boc - t-Bu -C6H11 O5 + 4H⁺)/4: 738.0]. The Boc groups of Peak-1 and Peak-2 were removed (each compound individually) by treating with TFA for about an 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Preparative HPLC with ACN/water (0.1 % TFA). The desired material (either Compound 61 or Compound 62) was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ion was found as [(M - C6H11 O5 + 5H⁺)/5: 465.1 ] for both Compound 61 and Compound 62. The stereochemistry of the trans- cyclopropyl dicarboxylate was arbitrarily assigned.

Step a. Preparation of dibenzyl 2,2'-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]

ethoxy}ethyl)azanediyl]diacetate

Benzyl bromo-acetate (4.0g, 17.4 mmol) was added into the solution INT-1 . L-Rhamnose-PEG1 - NH2 (1 .75 g, 7 mmol) and DIPEA (3.6 mL, 30 mmol) in 60 mL DMF, the resulting solution was stirred at room temperature overnight. The reaction solution was concentrated and purified by flash

chromatography to provide the desired product. Yield of oil product, LC/MS, 548.2 [M+H]+.

Step b. Preparation of 2,2'-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]-ethoxy}ethyl)- azanediyl]diacetic acid

Dibenzyl 2,2'-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy] ethoxy}ethyl)azanediyl] diacetate (1 .0g, 2 mmol) was dissolved into 10 mL MeOH and 10 mL ethyl acetate, then 200 mg of 5% palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere overnight. The palladium charcoal was removed by filtration after completed the reaction by LCMS. The filtrate was concentrated and used next step without any purification. Yield of oil product, LC/MS, 368.2 [M+H]+ .

Step c. Preparation of per-Boc-Compound 63

Standard procedure A: To the solution of 2,2'-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]- ethoxy}ethyl)-azanediyl]diacetic acid (40 mg, 0.1 mmol), triethyl amine (0.07 mL, 0.5 mmol) and INT-18 (320 mg, 0.2 mmol) in 5 mL DMF was added HATU (78 mg, 0.2 mmol). The reaction was stirred for 1 hr and then the resulting solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield of product (per-Boc-Compound 63), 220 mg, 65% yield.

Step d. Preparation of Compound 63

Standard procedure B: Per-Boc-Compound 63 from the previous step was treated in 2 mL

DCM and 2 ml_ trifluoroacetic acid at room temperature, the solution was stirred 1 0 min, then concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using trifluoroacetic acid as the modifier. Yield 0.120 g, 67% yield, lon(s) found by LCMS: [M+2H]/2 = 1229.2, [M+3H]/3 = 819.8,

[M+4H]/4 = 615.1 , [M+5H]/5 =492.3.

Example 153: Synthesis of Compound 64

Step a. O-Alkylation of S-Hydroxy-I .S-Benzenedicarboxyiic Acid Dimethyl Ester and Removal of the Cbz Group

Benzyl (2-bromoethyl)carbamate ( .5 g, 6.0 mmol), 5-hydroxy-l ,3-benzenedicarboxylic acid dimethyl ester (1 .0 g, 4.8 mmol), and cesium carbonate (2.0 g, 6 mmol) were stirred in DMF (10 mL) at 60°C for 4 hours. The mixture was diluted with D! water (200 mL) and extracted into ethyl acetate (75 mL, 3x). The combined organic extracts were dried over sodium sulfate and concentrated. The residue was purified by Silica gel chromatography (hexanes/ethyi acetate) to afford 1 .4 g of the Cbz protected intermediate as a white solid. The Cbz-protected intermediate was stirred in methanol (100 mL) in the presence of 5% Pd/C (200 mg) under 1 atmosphere of hydrogen for 2 hours. The mixture was filtered through ceiiie and concentrated to afford 1 .1 g of the product as a white solid. Yield: 83 %. LC/MS [ +H] = 254.4. Step b. Synthesis of the Hemi-succinate

Succinic anhydride (0.6 g, 6 mmol) and the Step-a product (1 .1 g, 4.3 mmol) were stirred in methanol (20 mL) for 2 hours then concentrated. The mixture was acidified with glacial acetic acid and concentrated. The residue was purified by RPLC (50 g, 5 % to 95% acetonitrile and water using 0.1 % TFA modifier). The pure fractions were pooled and lyophilized to afford 1 g of the product as a white solid. Yield: 67%. LC/MS [M+H]+ = 354.0.

Step c. Coupling to INT-1

HATU (0.75 g, 2 mmol) in DMF (2 mL) was added, dropwise over a period of 20 minutes, to a stirring mixture of the Step-b product (0.63 g, 1 .8 mmol), INT-1 (0.45g, 1 .8 mmol), and triethylamine (0.36 g, 3.6 mmol) in DMF (4 mL) and then stirred for an additional 20 minutes. The mixture was applied to RPLC (150 g, 5 % to 95% acetonitrile and water using 0.1 % TFA modifier). The pure fractions were pooled and lyophilized and then stirred for 30 minutes in a 1 /1 /2 mixture of THF/methanol/DI water containing LiOH (1 50 mg, 6 mmol). The mixture was acidified with glacial acetic acid and concentrated. The residue was purified by RPLC (50 g, 1 0 % to 95% acetonitrile and water using 0.1 % TFA modifier). The pure fractions were pooled and lyophilized to afford 625 mg of the product. Yield: 61 %. LC/MS

[M+H]+ = 559.6.

Step d. Synthesis of Compound 64

HATU (42 mg, 0.1 1 mmol), in DMF (1 mL) was added, dropwise over 20 minutes, to a stirring mixture of INT-18 (192, mg, 0.1 1 mmol), the Step-c product (30 mg, 0.054 mmol), and triethylamine (32 mg, 0.31 mmol) in DMF (2 mL). The reaction was stirred for 30 additional minutes and applied to RPLC (50 g, 30 % to 95% methanol and water using 0.1 % TFA modifier). The pure fractions were concentrated and then stirred in a 1 /1 mixture of TFA/DCM (5 mL) at ambient temperature for 20 minutes. The solvent was removed by rotovap and the residue was purified by RPLC (50 g, 0 % to 75% acetonitrile and water using 0.1 % TFA modifier). The pure fraction were pooled and lyophilized to afford the product as a white solid. Yield: 67%, 2 steps. LC/MS [M+3H/3] = 977.9. Example 154: Synthesis of Compound 65

Step a. Synthesis of the Dipeptide

HATU (2.89 g, 7.50mmol), in DMF (4 mL) was added, dropwise over 30 minutes, to a stirring mixture of Cbz-S-1 -amino-octanoic acid (2.00 g, 6.82 mmol), L-DAB-methyl ester hydrochloride (1 .75 g, 7.50 mmol), and triethylamine (2.07 mg, 20.5 mmol) in DMF (6 mL) and the reaction was stirred for an additional 30 minutes. The mixture was applied directly to RPLC (150 g, 15 % to 95% acetonitrile and water using 0.1 % TFA modifier). The pure fractions were pooled and lyophilized and then taken up in methanol (150 mL), 5% Pd/C (400 mg) was added and the mixture was stirred under an atmosphere of hydrogen for 12 hours. The mixture was filtered through Celite and concentrated to afford 2.2 g of the product as a white solid. Yield: 86%. LC/MS [M+H]+ = 374.6.

Step b. Coupling to 2,2'-{[(benzyloxy)carbonyl]azanediyl}diacetic Acid

The Step-b product was prepared from 2,2'-{[(benzyloxy)carbonyl]azanediyl}diacetic acid and the Step-a product in a similar procedure as described in Step a. Yield: 86%. LC/MS [M+H]+ = 844.6 .

Step c. Coupling to INT-2 and Methyl Ester Hydrolysis

HATU (406 mg, 1 .07 mmol), in DMF (2 mL) was added, dropwise over 30 minutes, to a stirring mixture of the Step-b product (820 mg, 0.97 mmol), INT-2 (375 mg, 1 .07 mmol), and triethylamine (294 mg, 2.91 mmol) in DMF (3 mL) and the reaction was stirred for an additional 30 minutes. The mixture was applied directly to RPLC (150 g, 15 % to 95% acetonitrile and water using 0.1 % TFA modifier). The pure fractions were pooled and lyophilized and then stirred for 30 minutes in a 1 /1 /2 mixture of

THF/methanol/DI water containing LiOH (1 16 mg, 4.86 mmol). The mixture was acidified with glacial acetic acid and concentrated. The residue was purified by RPLC (150 g, 10 % to 95% acetonitrile and water using 0.1 % TFA modifier). The pure fractions were pooled and lyophilized to afford 350 mg of the product. Yield: 86%. LC/MS [M-2boc+2H/2]+ = 475.4.

Step d. Preparation of Compound 65

The title compound was prepared from INT-63 and the Step-c product in a similar manner as described in the procedure for the preparation of Compound 64. Yield: 39%. LC/MS [M+3H/3]+ = 938.5.

The title compound was prepared analogously using standard procedure A and B of Example 152. lon(s) found by LCMS [M+3H/3] = 913.9, [M+4H/4] =685.6, [M+5H/5] =548.7. [M+6H/6] =457.4.

Example 156: Synthesis of Compound 67

The title compound was prepared analogously to Example 37, where INT-20 was prepared with D-aminooctanoic acid in place of L-aminooctanoic acid in that sequence of reactions, lon(s) found by LCMS: (M+3H)/3 = 946.9, (M+4H)/4 = 710.4, (M+5H)/5 = 568.5, (M+6H)/6 = 473.9

Example 157: Synthesis of Compound 68

Step a. Preparation of diethyl 2,2'-{[2-({2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethyl}amino)-2- oxoethyl]azanediyl}diacetate

To the solution of [bis(2-ethoxy-2-oxoethyl)amino]acetic acid (0.41 g, 2 mmol), 2-aminoethyl 6- deoxy-alpha-L-mannopyranoside (0.5 g, 2 mmol) in DMF (20 mL) was added EDC (400 mg, 2 mmol), HOBT (300 mg, 2 mmol), Hunig's base (0.42 mL, 3 mmol) at room temperature. The solution was stirred overnight. The resultant solution was removed DMF and purified by reversed phase liquid

chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. LC/MS 437.2 [M+H]

Step b. Preparation of 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]- 2-oxoethyl}azanediyl)diacetic acid

Lithium hydroxide (120 mg, 5mmol) in 5 mL H2O was added to the solution of diethyl 2,2'-({2-[(2- {2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)-diacetate (0.41 g, 0.94 mmol) in a mixture of 5 mL H2O, 5 mL MeOH and 5 mL THF. The resultant solution was stirred for 1 hour at room temperature. The solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoro-acetic acid as the modifier. The product was obtained as an oil. LC/MS 381 .1 [M+H]⁺

Step c. Preparation of Compound 68

The title compound was prepared analogously using standard procedure A and B of Example 152. lon(s) found by LCMS [M+3H/3] = 91 8.2, [M+4H/4] =688.9, [M+5H/5] =551 .3. [M+6H/6] =459.6.

Example 158: Synthesis of Compound 69 and Compound 70

The title compounds were prepared analogously to Example 157, where the 2,2'-({2-[(2-{2-[(6- deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)diacetic acid was replaced with (1 R, 2R)-3-{(1 E)-3-[(2-{2-[(6-deoxy-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-3-oxoprop-1 -en-1 - yl}cyclopropane-1 ,2-dicarboxylic acid (INT-49) in the first step of that sequence. The product was obtained as a separable mixture of diastereomers in the final step (RPLC), one diastereomer with a shorter retention time (more polar diastereomer) and one diastereomer with a longer retention time (less polar diastereomer). The relative stereochemistry of the diastereomers was arbitrarily assigned, lon(s) found by LCMS (for both diastereomers): (M+2H)/2 = 1403.3, (M+3H)/3 = 935.9, (M+4H)/4 = 702.2.

The title compound was prepared analogously to the preparation of Compound 68. lon(s) found by LCMS

[M+3H/3] = 823.1 , [M+4H/4] =618.3, [M+5H/5] =494.9. [M+6H/6] =412.6.

Step a. Synthesis of D-Asn-INT-11

A solution of Z-D-Asn-OH (87.9 mg, 0.33 mmol) and HOBT hydrate (70.7 mg, 0.463 mmol) in anhydrous DMF (1 mL) was cooled in an ice-water bath. EDC-HCI (76.8 mg, 0.4 mmol) was added, and the resulting mixture was stirred for 15 minutes. It was then added with a solution of INT-1 1 (319 mg, 0.3 mmol) in anhydrous DMF (1 mL) and DIPEA (130 mg, 1 mmol). After the reaction mixture was stirred for 1 hour, it was directly purified through C18 RPLC (50 g, 10 to 90 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (LCMS: [(M - 2Boc + 2H)/2]⁺ = 555). The material was re-dissolved in MeOH (~ 15 mL), and the solution was added with Pd/C. The resulting mixture was stirred under hydrogen for 4 hours. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation to dryness. Yield 1 85.2 mg, 52.5 %. Ion found by LCMS: [(M - Boc + 2H)/2]⁺ = 538.

Step b. Synthesis of octa-Boc-Compound 72 precursor

A mixture of INT-40 (59.2 mg, 0.0438 mmol) and D-Asn-INT-1 1 (105.2 mg, 0.0894 mmol) was dissolved in in anhydrous DMF (0.5 mL) and DIPEA (52 mg, 0.4 mmol). After the solution was cooled in an ice-water bath, it was added with a solution of HATU (36.6 mg, 0.0964 mmol) in DMF (0.5 mL) via syringe pump at a rate of 0.5 ml/hr. The reaction was stirred for 30 more minutes. It was then purified by RPLC (50 g, 20 to 100 % MeOH and water). Yield 72 mg, 42.7 %. Ions found by LCMS: [(M + 3H)/3]⁺ = 1224, [(M - Boc + 3H)/3]⁺= 1 190, [(M - 2Boc + 3H)/3]⁺= 1 156, [(M - 3Boc + 3H)/3]⁺= 1 123, [(M - 6Boc + 3H)/3]⁺= 1023.

Step c. Removal of the Boc groups

The step-b product (72 mg, 0.01 86 mmol) was dissolved in TFA (~ 0.5 mL) and stirred at room temperature for 15 minutes. It was then directly purified by RPLC (50 g, 5 to 38 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 27 mg, 35.5 %. Ions found by LCMS: [(M + 3H)/3]⁺ = 956.2, [(M + 4H)/4]⁺ = 71 7.3, [(M + 5H)/5]⁺ =574 , [(M + 6H)/6]⁺ = 478.5. Example 161 : Synthesis of Compound 73

Note: The absolute configuration of the trans-cyclopropane- 1 ,2-dicarboxamide is arbitrarily assigned Step a. Synthesis of 1 ,1-dibenzyl 2,3-di-tert-butyl (2S,3S)-cyclopropane-1 ,1 ,2,3-tetracarboxylate

To a solution of triphenylarsine (306.2 mg, 1 mmol) in EtOH at 0 °C was added di-benzyl malonate (582.8 mg, 2.05 mmol) in EtOH (2 mL). The mixture was added drop-wise a solution of di-tert- butyl acetylene dicarboxylate (452.6 mg, 2 mmol) in acetonitrile (1 .4 mL) via syringe pump at a rate of 1 .4 ml/hr. After the addition, the reaction was stirred at 0 °C to room temperature for 5 days, then concentrated by rotary evaporation. The residue was purified through silica gel column chromatography (80 g, 0 to 25 % EtOAc in hexane). The collected fractions were concentrated by rotary evaporation and further dried in high vacuum to afford the product as a clear oil (674.2 mg, 66.0 %). Ion found by LCMS: [M - 2tBu + H]⁺ = 399.

Step b. Synthesis of (1S,2S)-3,3-bis[(benzyloxy)carbonyl]cyclopropane-1 ,2-dicarboxylic acid

The step-a product (624.2 mg, 1 .22 mmol) was dissolved in DCM (~ 3 mL), and the solution was treated with a mixture of TFA/anisole (2:0.2 mL). After the mixture was stirred at room temperature overnight, it was concentrated by rotary evaporation. The residue was purified by RPLC (100 g, 1 0 to 65 % acetonitrile and water). Yield 434.5 mg of thick oil, 89.5 %. Ion found by LCMS: [M + H]⁺ = 399.

Step c. Synthesis of di-carboxylic acid-Compound 73 precursor

A mixture of INT-20 (151 mg, 0.886 mmol) and the step-b product (17.3 mg, 0.0434 mmol) was dissolved in anhydrous DMF (0.5 mL) and DIPEA (65 mg, 0.5 mmol). After the solution was cooled in an ice-water bath, it was drop-wise added with a solution of HATU (37 mg, 0.1 mmol) in anhydrous DMF (0.5 mL) via syringe pump at a rate of 1 ml/hr. After the addition, the reaction was stirred for 30 more minutes and directly purified by RPLC (50 g, 30 to 100 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid. The material was re-dissolved in MeOH (~ 15 mL) and added with Pd/C. The mixture was stirred under hydrogen overnight. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried in high vacuum to a white solid. Yield

96.2 mg, 61 .8 % over two steps. Ions found by LCMS: [(M + 3H)/3]⁺ = 1 1 99, [(M - 4Boc + 3H)/3]⁺ = 1025.

Step d. Synthesis of Rhamnose Compound 73 precursor

The step-c product (96.2 mg, 0.0268 mmol) was dissolved in anhydrous DMF (0.5 mL) and DIPEA (10.3 mg, 0.1 mmol). After the solution was cooled in an ice-water bath, it was added with HATU (10.2 mg, 0.0268 mmol). The mixture was stirred for 15 minutes, then INT-1 (1 1 mg, 0.044 mmol) was added. The resulting mixture was stirred at 0 °C to room temperature overnight. It was directly purified by RPLC (50 g, 20 to 100 % MeOH and water). Yield 57.6 mg, 56.2 %. Ions found by LCMS: [(M + 3H)/3]⁺ = 1275, [(M - 3Boc + 3H)/3]⁺ = 1 176, [M - 4Boc + 3H)/3]⁺ = 1 142, [(M - 5Boc + 3H)/3]⁺ = 1 109.

Step e. Removal of the Boc group

The step-d product (57.6 mg, 0.0151 mmol) was dissolved in TFA (~ 1 mL). The solution was stirred for 15 minutes and then directly purified through HPLC (5 to 23 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 13.1 mg, 21 .9 %. Ions found by LCMS: [(M + 4H)/4]⁺ = 706, [(M + 5H)/5]⁺ = 565, [(M + 6H)/6]⁺ = 471 . xample 162: Synthesis of Compound 74

The title compound was prepared analogously to Example 152, where 2,2'-({2-[(2-{2-[(6-deoxy- alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)diacetic acid was replaced with (1 R, 2R)-3-({4-[(2-{2-[(6-deoxy-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanamido}methyl)cyclopropane-1 ,2-dicarboxylic acid (INT-46) in the first step of that sequence. lon(s) found by LCMS: (M+2H)/2 = 1432.8, (M+3H)/3 = 955.6, (M+4H)/4 = 716.9.

The dicarboxylic acid INT-46 (0.052 g, 0. 106 mmol) and INT-48 (0.266 g, 0.170 mmol) were dissolved in 5 mL of DMF followed by addition of DI PEA (0.046 mL, 0.26 mmol). To this mixture was added HATU (0.100 g, 0.264 mmol) in 1 .5 mL of DMF via syringe pump in 2 hours at room temperature. After completion of HATU addition, the reaction mixture was stirred for 1 hr at room temperature. Then the reaction mixture was concentrated under reduced pressure. The residue was purified by reversed phase prep-HPLC with ACN/water (0.1 %TFA) to give the per-Boc-Compound 75 as Peak-1 (polar) and per-Boc-Compound 76 as Peak-2 (less polar). The mass spectrum showed strong detectable positive charge signals at tr = 2.86 and 3.07 min with 7 min (60-95%) method [found positive ion (M - 3Boc +

3H⁺)/3: 1056.2]. The Boc groups of Peak-1 and Peak-2 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Preparative HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M + 5H⁺)/5: 534.0, (M + 4H⁺)/4: 667.0] at tr = 2.21 min with 5 min (5-95%) method for Compound 75 [(M + 5H⁺)/5: 534.0, (M + 4H⁺)/4: 667.2] at tr = 2.41 min with 5 min (5-95%) method for Compound 76. The stereochemistry of the trans-cyclopropane dicarboxamide was arbitrarily assigned.

Example 164: Synthesis of Compound 77 and Compound 78

The dicarboxylic acid INT-49 (0.0599 g, 0. 138 mmol) and INT-48 (0.416 g, 0.276 mmol) were dissolved in 5 mL of DMF followed by addition of DI PEA (0.060 mL, 0.35 mmol). To this mixture was added HATU (0.131 g, 0.345 mmol) in 1 .5 mL of DMF via syringe pump in 2 hours at room temperature. After completion of HATU addition, the reaction mixture was stirred for 1 hr at room temperature. Then the reaction mixture was concentrated under reduced pressure. The residue was purified by reversed phase prep-HPLC with ACN/water (0.1 %TFA) to give the per-Boc-Compound 77 as Peak-1 (polar) and per-Boc-Compound 78 as Peak-2 (less polar). The mass spectrum showed strong detectable positive charge signals at tr = 2.96 and 3.18 min with 7 min (60-95%) method [found positive ion (M - CeHnCH -

4Boc + 5H⁺)/5: 571 .8]. The Boc groups of Peak-1 and Peak-2 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Preparative HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M + 5H⁺)/5: 522.4, (M + 4H⁺)/4: 652.4, (M + 3H⁺)/3: 869.4] at tr = 2.13 min with 5 min (5-95%) method for Compound 77 [(M + 5H⁺)/5: 522.0, (M + 4H⁺)/4: 652.2, (M + 3H⁺)/3: 869.6] at tr = 2.34 min with 5 min (5-95%) method for Compound 78. The stereochemistry of the trans-cyclopropane dicarboxamide was arbitrarily assigned.

The dicarboxylic acid INT-46 (0.0340 g, 0. 0690 mmol) and INT-50 (0.235 g, 0.138 mmol) were dissolved in 5 mL of DMF followed by addition of DI PEA (0.030 mL, 0.17 mmol). To this mixture was added HATU (0.656 g, 0.172 mmol) in 1 .0 mL of DMF via syringe pump in 2 hours at room temperature. After completion of HATU addition, the reaction mixture was stirred for 1 hr at room temperature. Then the reaction mixture was concentrated under reduced pressure. The residue was purified by reversed phase prep-HPLC with ACN/water (0.1 %TFA) to give the per-Boc-Compound 79 as Peak-1 (polar) and per-Boc-Compound 80 as Peak-2 (less polar). The mass spectrum showed strong detectable positive charge signals at tr = 3.07 and 3.26 min with 7 min (60-95%) method [found positive ion (M - ΟβΗηΟδ - 4Boc + 4H⁺)/4: 852.6, (M - CeHnOs - Boc + 3H⁺)/3: 1200.8, (M - CeHnC - Boc + 3H⁺)/3: 1207.0]. The Boc groups of Peak-1 and Peak-2 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Preparative HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M + 6H⁺)/6: 478.6, (M + 5H⁺)/5: 574.8] at tr = 2.03 min with 5 min (5-95%) method for Compound 79, [(M + 6H⁺)/6: 478.4, (M + 5H⁺)/5: 574.8] at tr = 2.14 min with 5 min (5-95%) method for Compound 80. The stereochemistry of the trans- cyclopropane dicarboxamide was arbitrarily assigned.

Example 166: Synthesis of Compound 81

Part 1. Preparation of the Asparagine Decapeptide

Step a. Synthesis of Z-Thr-Asn-tert-butyl ester

A mixture of Z-Thr-OH (2.79 g, 1 1 mmol) and HOBT (2.14 g, 14 mmol) in anhydrous DMF (8 ml_) was cooled in an ice-water bath and added with EDC HCI (2.3 g, 12 mmol). After stirred for 30 minutes, the reaction was added with a suspension of H-Asn-OtBu (1 .88 g, 1 0 mmol) and DIPEA (2.6 g, 20 mmol) in DMF (10 mL) and DCM (10 mL). The reaction was continued overnight and then extracted with water (50 mL) and DCM (40 mL x 2). The combined organic layers were concentrated by rotary evaporation. The residue was purified by RPLC (100 g, 5 to 50 % acetonitrile and water). Yield 3.5 g, 82.6 %. Ions found by LCMS: [M + H]⁺ = 424, [M - tBu + H]⁺ = 368. Step b. Synthesis of Z-Thr-Asn-OH

The step-a product (3.5 g, 8.26 mmol) in DMC (5 mL) was added with TFA (10 mL) and thioanisole (3 mL). After the mixture was stirred at room temperature for 1 day, it was concentrated by rotary evaporation. The residue was purified by RPLC (150 g, 5 to 31 % acetonitrile and water). Yield 448.7 mg, 14.8 %. Ion found by LCMS: [M + H]⁺ = 368.

Step c. Synthesis of Z-Thr-Asn-INT-11

A mixture of the step-b product (2368 mg, 1 mmol) and HOBT (214.2 mg, 1 .4 mmol) in anhydrous DMF (1 mL) was cooled in an ice-water bath and added with EDC HCI (209 mg, 1 .1 mmol). After stirred for 30 minutes, the reaction was added with INT-1 1 (1 .06 g, 1 mmol) and DIPEA (325 mg, 2.5 mmol) in DMF (1 .5 mL). The reaction was continued for 2 hours and then purified by RPLC (100 g, 15 to 100 % MeOH and water, using 0.1 % TFA as modifier). Yield 1 .3 g, 92 %. Ions found by LCMS: [(M - 2Boc + 2H)/2]⁺ = 606.5. Step d. Removal of the Cbz protection

Step-c product (1 .3 g, 0.92 mmol) was dissolved in MeOH (30 mL). The solution was added with Pd/C and stirred under hydrogen for 5 hours. Pd/C was removed, and the filtrate was concentrated by rotary evaporation to dryness. Yield 1 .1 g, 93.6 %. Ion found by LCMS: [(M - Boc + 2H)/2]⁺ = 589. Step e. Synthesis of the Asparagine Decapeptide

A mixture of the step-d product (1 .1 g, 0.861 mmol), N^a-Z-N^Y-Boc-L-2,4-dimaminobutyric acid dicyclohexylammonium (533.7 mg, 1 mmol) and DIPEA (260 mg, 2 mmol) in anhydrous NMP (3 mL) was cooled in an ice-water bath and drop-wise added with a solution of HATU (400 mg, 1 .06 mmol) in NMP (1 .5 mL) via syringe pump at a rate of 1 ml/hr. After the addition, the resulting mixture was stirred for 1 hour and directly purified by RPLC (50 g, 20 to 100 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid. The material was re-dissolved in MeOH (30 mL) and added with Pd/C. The mixture was stirred under hydrogen for 4 hours. Pd/C was removed, and the filtrate was concentrated by rotary evaporation to dryness. Yield 601 mg, 47.2 %. Ion found by LCMS: [(M - Boc +2H)/2]⁺ = 689.

Step a. Synthesis of Octa-Boc Compound 81 precursor

A mixture of INT-66 (56 mg, 0.0723 mmol) and the asparagine decapeptide (235 mg, 0.159 mmol) was dissolved in anhydrous DMF (1 mL) and DIPEA (65 mg, 0.5 mmol). After cooled in an ice- water bath, the mixture was drop-wise added with a solution of HATU (70.4 mg, 0.185 mmol) in DMF (0.5 mL) at a rate of 1 ml/hr. After the addition, the reaction was stirred for 30 more minutes and then purified by RPLC (150 g, 10 to 100 % MeOH and water, using 0.1 % TFA as modifier). The collected fractions were concentrated by rotary evaporation to a white solid. Yield 68.5 mg, 25.7 %. Ions found by LCMS: [(M - Boc + 3H)/3]⁺ = 1 1 98, [(M - 2Boc + 3H)/3]⁺ = 1 165, [(M - 3Boc + 3H)/3]⁺ = 1 132, [(M - 4Boc + 3H)/3]⁺ = 1098.

Step b. Removal of the Boc groups to give Compound 81

The step-a product (68.5 mg, 0.186 mmol) was dissolved in TFA (~ 1 mL). The solution was stirred at room temperature for 15 minutes, then directly purified by RPLC (50 g, 0 to 33 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 24.5 mg, 34.7 %. Ions found by LCMS: [(M + 3H)/3]⁺ = 965, [(M + 4H)/4]⁺ = 724, [(M + 5H)/5]⁺ = 579, [(M + 6H)/6]⁺ = 483.

Step a. Preparation of 4,4'-{[1 ,4-dimethoxy-1 ,4-dioxobutane-2,3-diyl]diazanediyl}bis(4-oxobutanoic acid), meso-

Dimeihyl 2.3-diamino-succinate (meso) (176 mg, 1 mmo!) and succinic anhydride (600 mg, 6 mmo!) were dissolved into 5 mL 10% pyridine in DC , the resulting solution was stirred overnight and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 0% to 100% acetonitrile and water, using trifluoroacetic acid as the modifier. The product was obtained as an oil, LCMS: 377.1 , [M+H]+.

Step b. Preparation of dimethyl 2,3-(meso)-bis(4-oxo-4-{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyloxan-2-yl]oxy}ethoxy)ethyl]amino}butanamido)butanedioate 4,4'-{[1 ,4-Dimethoxy-1 ,4-dioxobutane-2,3-diyl]diazanediyl}bis(4-oxobutanoic acid), meso- (90 mg, 0.23 mmol) was dissolved into 5 mL DMF, then INT-1 (120 mg, 0.5 mmol), EDC (100 mg, 0.5 mmol), HOBT (80 mg, 0.5 mmol), Hunig's base (0.14 mL, 1 mmol) were added the solution at room temperature. The resultant solution was stirred overnight. The resultant solution was removed DMF and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. LC/MS 843.4 [M+H].

Step c. Synthesis of 2,3-(meso)-bis(4-oxo-4-{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyloxan-2-yl]oxy}ethoxy)ethyl]amino}butanamido)butanedioic acid

Lithium hydroxide (12 mg, 0.5 mmol) in 2 mL H2O was added to the solution of dimethyl 2,3- (meso)-bis(4-oxo-4-{[2-(2-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}ethoxy) - ethyl]amino}butanamido)butanedioate (30 mg, 0.035 mmol) in 2 mL MeOH and 2mL THF, and the mixture was stirred at room temperature for 1 hours at room temperature. The reaction solution was quenched by drops of acetic acid. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoro-acetic acid as the modifier. The product was obtained as an oil. LC/MS 815.3, [M+H]+.

Step d. Preparation of Compound 82.

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3] = 1062.9, [M+4H/4] =797.5, [M+5H/5]=638.2, [M+6H/6]=531 .9.

Step a. Preparation of diethyl 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)diacetate

To the solution of [bis(2-ethoxy-2-oxoethyl)amino]acetic acid (4.4 g, 18 mmol), INT-1 (4.51 g, 18 mmol) in DMF (50 mL) was added EDC (4 g, 20 mmol), HOBT (2.7 g, 20 mmol), Hunig's base (4.2 mL, 30 mmol) at room temperature. The solution was stirred overnight. The resultant solution was removed DMF and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. LC/MS 481 .2 [M+H]

Lithium hydroxide (600 mg, 25 mmol) in 10 mL H20 was added to the solution of diethyl 2,2'-({2- [(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)-diacetate (4.80 g, 10mmol) in mix solvent of 10 mL H20, 20 mL MeOH and 20 mL THF. The resultant solution was stirred for 1 hours at room temperature. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. LC/MS 425.2 [M+H]⁺

Step c. Preparation of Compound 83

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3] = 928.2, [M+4H/4] =696.4, [M+5H/5]=557.3, [M+6H/6]=464.6.

Step a. Synthesis of N-[(benzyloxy)carbonyl]-3-carbamimidamido-L-alanine

L-2-amino-3-guanidinopropionic acid (1 .83 g, 10 mmol) was dissolved in a solution of NaOH (800 mg, 10 mmol) in water (10 mL). After cooled in ice-water bath, the mixture was added with a solution of N-

(benzyloxycarbonyloxy)succinimide (2.73 g, 1 1 mmol) in THF (10 mL). The resulting mixture was stirred at 0 °C to room temperature overnight. THF was then partially removed by rotary evaporation, and the residue was extracted with a 1 :1 mixture of EtOAc/hexane (40 mL). The aqueous layer was purified by RPLC (150 g, 0 to 40 % acetonitrile and water). Yield 2.05 g, 71 .3 %. Ion found by LCMS: [M + H]⁺ = 281 . Step b. Synthesis of N-[(benzyloxy)carbonyl)]-3-carbamimidamido-Ala-INT-11

To a solution of a mixture of N-[(benzyloxy)carbonyl]-3-carbamimidamido-L-alanine (420 mg, 1 .5 mmol) and INT-1 1 (1 .06 g, 1 mmol) in anhydrous DMF (2 mL) was added DMAP (24.4 mg, 0.2 mmol) and HATU (570 mg, 1 .5 mmol). After the reaction was stirred overnight, it was added with DIPEA (130 mg, 1 mmol) and continued for 6 more hours. It was then directly purified through RPLC (100 g, 20 to 80 % MeOH and water). Yield 980 mg, 74 %. Ion found by LCMS: [(M - Boc + 2H)/2]⁺ = 613.

Step c. Synthesis of 3-carbamimidamido-Ala-INT-11

The step-b product (185.4 mg, 0.14 mmol) was re-dissolved in MeOH (20 mL), and the solution was added with Pd/C. The mixture was stirred under hydrogen for 3 hours. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation to dryness. Yield 165 mg, 98%. Ion found by LCMS: [(M - Boc + 2H)/2]⁺ = 546.

Step d. Synthesis of octa-Boc-Compound 84 precursor

A mixture of the step-c product (120 mg, 0.1 mmol), INT-40 (66.8 mg, 0.494 mmol), and DMAP (66.8 mg) was dissolved in anhydrous DMF (2 mL). The solution was drop-wise added with a HATU (45.6 mg, 0.12 mmol) solution in DMF (0.5 mL) via syringe pump at a rate of 1 ml/hr. After the addition, the resulting mixture was stirred for one more hour. It was then directly purified by RPLC (50 g, 20 to 95 % MeOH in water, using 0.1 % TFA as modifier). Yield 1 15 mg, 63 %. Ions found by LCMS: [(M - Boc + 3H)/3]⁺ = 1 1 99, [(M - 2Boc + 3H)/3]⁺ = 1 166, [(M - 3Boc + 3H)/3]⁺ = 1 132, [(M - 4Boc + 3H)/3]⁺ = 1099, [(M - 5Boc + 3H)/3]⁺ = 1066).

Step e. Removal of the Boc group

The step-d product was dissolved a 1 :1 mixture of TFA:DCM (~ 1 mL). The solution was stirred for 15 minutes, then directly purified through HPLC (0 to 22.4 % acetonitrile in water, using 0.1 % TFA as modifier). Yield 14.8 mg, 12.5 %. Ions found by LCMS: [(M + 3H)/3]⁺ = 966, [(M + 4H)/4]⁺ = 724, [(M + 5H)/5]⁺ = 580, [(M + 6H)/6]⁺ = 483.

Step a. Synthesis of benzyl (3R,4R)-3,4-bis{[(2S)-1-methoxy-1-oxooctan-2- yl]carbamoyl}pyrrolidine-1-carboxylate

A solution of transl -[(benzyloxy)carbonyl]pyrrolidine-3,4-dicarboxylic acid (3.69g, 12.58 mmol) and methyl (2S)-2-aminooctanoate (4.58g, 26.42 mmol), DIEA(13.8ml_, 79.3mmol) in 20ml_ of DMF was treated with HATU (10. Og, 26.4mmol). After stirring for 30 minutes at room temperature the reaction was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using formic acid (0.1 %) as the modifier. Yield 1 .44 g, 19%. lon(s) found by LCMS: M+H = 604.4

Step b. Synthesis of (2S,2'S)-2,2'-[{(3R,4R)-1-[(benzyloxy)carbonyl]pyrrolidine-3,4- diyl}bis(carbonylazanediyl)]dioctanoic acid

A solution of benzyl (3R, 4R)-3,4-bis{[(2S)-1 -methoxy-1 -oxooctan-2-yl]carbamoyl}pyrrolidine-1 - carboxylate (1 .44g, 2.39mmol) dissolved in THF (1 OmL) was treated with a solution of LiOH(0.143g,

5.96mmol) dissolved in water (10ml_). After stirring for 30 minutes all starting material was consumed by LCMS. Work-up: the reaction was made slightly acidic with concentrated HCI, extracted into ethyl acetate, and dried over sodium sulfate. The resulting crude product was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using formic acid (0.1 %) as the modifier. Yield 1 .14 g, 83%.

Step c. Synthesis of Cbz-pyrrolidine-polymyxin B dimer

A solution of racemic (2S,2'S)-2,2'-[{(3R,4R)-1 -[(benzyloxy)carbonyl]pyrrolidine-3,4- diyl}bis(carbonylazanediyl)]dioctanoic acid (0.401 g, 0.697mmol), INT-18 (2.18g, 1 .39mmol), DIEA(0.76ml_, 4.39mmol) dissolved in DMF(5ml_) was treated with a solution of HATU(0.557g, 1 .46mmol) dissolved in DMF(2mL), dropwise over 60 minutes. The reaction was stirred for an additional hour, then taken-on to the next step without further purification. Step d. Synthesis of Pyrrolidine-polymyxinB dimer

A solution of crude Cbz-pyrrolidine-polymyxinB dimer in DMF was treated with 5%Pd/C (0.75g) and vacuum flushed with hydrogen and stirred under a hydrogen atmosphere. LCMS after 2h shows complete conversion. The reaction was filtered through Celite, concentrated, and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using no modifier. Yield 1 .39 g, 56%. lon(s) found by LCMS: [(M- 2Boc)+3H]/3 = 1 1 1 1 .4, [(M-3Boc)+3H]/3 = 1078.0, [(M-4Boc)+3H]/3 = 1044.7.

Step e. Synthesis of Rhamnose Pyrrolidine-polymyxinB dimer

A solution of pyrrolidine-polymyxinB dimer (0.300g, 0.085mmol), INT-2 (0.0298g, 0.085mmol), and DIEA (0.049mL, 0.280mmol) was treated with a solution of HATU(0.036g, 0.0934mmol) in

DMF(1 .OmL), dropwise over 60 minutes. After an additional hour stirring the solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using 0.1 % TFA as the modifier. Yield 0.41 6 g, 127% (not pure), lon(s) found by LCMS: Did not ionize.

Step f. Deprotection to Compound 85

A solution of rhamnose pyrrolidine-polymyxinB dimer (0.328g, 0.085mmol) dissolved in

DCM(2mL) was treated with TFA(2mL) for 5 minutes. The solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 1 00% acetonitrile and water, using 0.1 % formic acid as the modifier. Yield 0.1 90 g, 67%. lon(s) found by LCMS: (M+3H)/3 = 955.8, (M+4H)/4 = 716.9, (M+5H)/5 = 573.7.

Example 171 : Synthesis of Compound 86

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3] = 1029.9, [M+4H/4] =772.7, [M+5H/5]=61 8.4, [M+6H/6]=515.5.

Step a. Synthesis of diethyl 1-{4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl) amino]-4-oxobutanoyl}pyrrolidine-2,5-dicarboxylate

To the solution of diethyl pyrrolidine-2,5-dicarboxylate (1 .0 g, 4.6 mmol), INT-2 (4-[(2-{2-[(6- deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid,1 .6 g, 4.6 mmol) in DMF (20 mL) was added EDC (1 g, 5 mmol), HOBT (800 mg, 5 mmol), Hunig's base (1 .4 ml_, 10 mmol) at room temperature. The solution was stirred overnight. The resultant solution was removed DMF and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. LC/MS 549.3 [M+H]

Step b. Synthesis of 1-{4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl) amino]-4- oxobutanoyl}pyrrolidine-2,5-dicarboxylic acid

Lithium hydroxide (120 mg, 5 mmol) in 5 mL H20 was added to the solution of diethyl 1 -{4-[(2-{2- [(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl) amino]-4-oxobutanoyl}pyrrolidine-2,5-dicarboxylate (1 .0 g, 1 .8 mmol) in mix solvent of 5 mL MeOH and 5 mL THF. The resultant solution was stirred for 1 hours at room temperature. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoro-acetic acid as the modifier. The product was obtained as an oil. LC/MS 493.2 [M+H]⁺

Step c. Preparation of Compound 87

The title compound was prepared in an analogous manner as described in standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3] = 955.6, [M+4H/4] =71 6.9, [M+5H/5]=573.7, [M+6H/6]=478.3.

xample 173: Synthesis of Compound 88

Step a. Coupling of L-Nor-Leu Methyl Ester to racemic-trans-1-(tert-butoxycarbonyl)pyrrolidine- 3,4-dicarboxylic acid

To a solution of racemic-trans-1 -(tert-butoxycarbonyl)pyrrolidine-3,4-dicarboxylic acid (257.5 mg, 0.99 mmol), L-Norleu-OMe-hydrochloride (375.29 mg, 2.07 mmol), and diisopropylethylamine (384.79 mg, 2.98 mmol) in DMF (10 ml_) was added a solution of HATU (0.29 g, 0.77 mmol) in DMF (2 ml_) via a syringe pump at a rate of 2.0 mL/hr. After the addition, the reaction mixture was stirred for an additional 1 hour at room temperature. The reaction mixture was purified by reversed phase liquid chromatography

(RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 50% acetonitrile and water, using 0.1 % TFA as modifier to yield two diastereomers: a more polar diastereomer (Step-a more polar isomer) (1 13.1 mg, 22%) and a less polar diastereomer (Step-a less polar isomer) (92.9 mg, 17%). LC/MS [(M- Boc)+H]+ = 414.2 (artefactual loss of 1 boc group) for both boc-protected intermediates (methyl 2- [((3R,4R)-4-{N-[(1 S)-1 -(methoxycarbonyl)pentyl]carbamoyl}-1 [(tert-butyl)oxycarbonyl]pyrrolidin-3- yl)carbonylamino](2S). Note: the isomers were taken forward separately in subsequent syntheses. Step b. Synthesis of INT-37a

A solution of the Step-a more polar isomer (379 mg, 1 .46 mmol) was stirred in TFA:CH2Cl2 (1 :2, 9 mL) at room temperature until completion. The volatiles were removed under reduced pressure then purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 0 to 75% acetonitrile and water, using 0.1 % TFA as modifier to yield INT-37a (217.9 mg, 36%). Ion found by LC/MS [M+H]+ = 414.2.

Step c. Conjugation of INT-37a with INT-2

INT-37a (217.3 mg, 0.41 mmol), INT-2, (4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanoic acid, 159.2 mg, 0.45 mmol), HOBt (61 .22 mg, 0.45 mmol) and triethylamine (129.21 mg, 1 .28 mmol) were stirred in DMF (3 mL). To this solution was added a prepared solution of EDCI (70.34 mg, 0.45 mmol) in DMF (2 mL). The reaction was stirred at room temperature until completion. The reaction mixture was diluted with ethyl acetate and washed with 1 N HCI aqueous solution and saturated NaHCCb aqueous solution. The organic layer was dried over Na2S04 and filtered then concentrated. The residue was purified by reversed phase liquid

chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10 to 50% acetonitrile and water, using 0.1 % TFA as modifier to yield the desired compound (204.2 mg, 66%). Ion found by LC/MS [M+H]+ = 747.3.

Step d. Synthesis of INT-38a

To a solution of Step-c product (204.2 mg, 0.27 mmol) in MeOH:THF:H₂0 (1 :1 :2, 8 mL) at room temperature was added LiOH (14 mg, 0.58 mmol) until completion monitoring by analytical HPLC. After the reaction was completed, it was quenched with 1 N HCI aqueous solution to pH 3 then concentrated under reduced pressure. The residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0 to 50% acetonitrile and water, using 0.1 % TFA as modifier to yield the desired compound (65.2 mg, 33%). Ion found by LC/MS [M-H]+ = 717.3.

Step e. Synthesis of Compound 88

A solution of HATU (45.70 mg, 0.12 mmol) in DMF (0.3 mL) was added via a syringe pump at a rate of 2.5 mL/hr into a solution of INT-38a (40.0 mg, 0.056 mmol), INT-17 (183.89 mg, 0.12 mmol) and diisopropylethylamine (43.12 mg, 0.33 mmol) in DMF (0.9 mL). The reaction mixture was stirred for an additional hour then purified reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 30 to 100% methanol and water, using 0.1 % TFA as modifier to yield the Boc-protected intermediate. The Boc-protected intermediate was stirred in TFA:DCM (1 :1 , 4 mL) at room temperature for 20 minutes. After the reaction was completed, the reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0 to 40% acetonitrile and water, using 0.1 % TFA as modifier to yield the title compound (1 19.5 mg, 55%). lon(s) found by LC/MS [(M+3H)/3]+ = 914.0, [(M+4H)/4]+ = 685.5, [(M+5H)/5]+ = 549.3 and [(M+6H)/6]+ = 457.9.

Example 174: Synthesis of Compound 89

The dimethyl ester INT-51 (0.102 g, 0. 137 mmol) was dissolved in 6 mL of THF/H2O (1 :1 ) followed by treating with LiOH (0.0069 g, 0.29 mmol). After 30 minutes, the reaction was finished and acidified with 1 N HCI and extracted with EtOAc to give the diacid without purification for next step. The diacid and INT-18 (0.4277 g, 0.2735 mmol) were dissolved in 5 mL of DMF followed by addition of DIPEA (0.071 mL, 0.41 mmol). To this mixture was added HATU (0.130 g, 0.342 mmol) in 2.0 mL of DMF via syringe pump in 2 hours at room temperature. After completion of HATU addition, the reaction mixture was stirred for 1 hr at room temperature. Then the reaction mixture was concentrated under reduced pressure. The residue was purified by reversed phase prep-HPLC with ACN/water (0.1 %TFA) to give the per-Boc-Compound 89. The mass spectrum showed strong detectable positive charge signals at tr = 3.77 min with 7 min (60-95%) method [found positive ion (M -CeHnOs - 6Boc + 6H⁺)/6: 507.4, (M - CeH C - 6Boc + 8H⁺)/8: 408.2]. The Boc groups of Per-Boc-Compound 89 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Prep-HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M + 6H⁺)/6: 469.2, (M + 5H⁺)/5: 562.8] at tr = 2.03 min with 5 min (5-95%) method for Compound 89. Example 175: Synthesis of Compound 90

The dimethyl ester INT-52 (0.133 g, 0. 178 mmol) was dissolved in 8 mL of THF/H2O (1 :1 ) followed by treating with LiOH (0.0090 g, 0.37 mmol). After 30 minutes, the reaction was finished and acidified with 1 N HCI and extracted with EtOAc to give the diacid without purification for next step. The diacid and INT-18 (0.5577 g, 0.3566 mmol) were dissolved in 5 mL of DMF followed by addition of DIPEA (0.093 mL, 0.53 mmol). To this mixture was added HATU (0.1695 g, 0.4577 mmol) in 2.0 mL of DMF via syringe pump in 2 hours at room temperature. After completion of HATU addition, the reaction mixture was stirred for 1 hr at room temperature. Then the reaction mixture was concentrated under reduced pressure. The residue was purified by reversed phase C18 with ACN/water (0.1 %TFA) to give the per- Boc-Compound 90. The mass spectrum showed a strong detectable positive charge signal at tr = 4.26 min with 7 min (60-95%) method [found positive ion (M - 3Boc + 3H⁺)/3: 1 170.4]. The Boc groups of Per- Boc Compound 90 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a prep-PLC with

ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M + 6H⁺)/6: 468.8, (M + 5H⁺)/5: 562.2, (M + 4H⁺)/4: 702.4] at tr = 2.23 min with 5 min (5-95%) method for Compound 90.

Example 176: Synthesis of Compound 91

INT-56 (0.224 g, 0.149 mmol) was dissolved in 6 mL of THF/water (1 :1 ) and treated with LiOH (0.0075 g, 0.312 mmol). After acidification THF was removed and 3mL of NMP was added. Then most of water was removed and the residue was load to pep-HPLC to purify. 0.219 g of INT-56 diacid was obtained (100%). Positive mass ions were found as (M - 2 CeHnC + 2H⁺)/2: 593.3, (M - CeHnC + 2H⁺)/2: 666.2, (M + 2H⁺)/2: 739.2, tr = 3.93 minutes with 8 min (5-95%) method.

The diacid, INT-18 (0.465 g, 0.297 mmol) and DIPEA (0.078 mL, 0.45 mmol) were mixed in 5 mL of DMF. HATU (0.141 g, 0.371 mmol) was added via syringe pump (2 mL, 1 ml/hr). The reaction solution was stirred for another hour after addition. DMF was removed and the residue was purified by prep-HPLC to give per-Boc- Compound 91 , 0.323 g, 48%. Positive mass ions were found as (M - 2C6H11 O4 + 4H⁺)/4: 1067.6, (M -4Boc - CeHn Ot + 4H⁺)/4: 1005.4, tr = 2.55 minutes with 7 min (60-95%) method. The Boc groups of Per-Boc- Compound 91 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto prep- HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M - 2C6H11 O4 + 5H⁺)/5: 655.8, (M - C6H11 O4 + 5H⁺)/5: 684.8, (M + 5H⁺)/5: 714.2, (M + 4H⁺)/4: 892.5] at tr = 2.02 min with 5 min (5-95%) method for Compound 91 .

INT-57 (0.069 g, 0.090 mmol), INT-18 (0.282 g, 0.180 mmol) and DIPEA (0.047 ml_, 0.27 mmol) were mixed in 5 mL of DMF. HATU (0.0857 g, 0.226 mmol) was added via syringe pump (2 ml_, 1 ml/hr). The reaction solution was stirred for another hour after addition. DMF was removed and the residue was purified by prep-HPLC to give per-Boc-Compound 92, 0.193 g, 55%. Positive mass ions were found as (M - 3Boc+ 4H⁺)/4: 889.6, (M - 3Boc+ 3H⁺)/3: 1 186.8, (M - 3Boc - CeHnC + 4H⁺)/4: 850.6, tr = 3.08 minutes with 7 min (60-95%) method. The Boc groups of Per-Boc-Compound 92 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto prep- HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M - C₆Hi i0₄S + 6H⁺)/6: 443.0, (M - CeH C S + 5H⁺)/5: 531 .4, (M + 6H⁺)/6: 476.6, (M + 5H⁺)/: 571 .4] for Compound 92.

Step a. Preparation of 4-benzyl 1-ferf-butyl W-{[(2S)-1-methoxy-1-oxooctan-2-yl]carbamoyl}-L- aspartate

Methyl (2S)-2-isocyanato-octanoate (200 mg, 1 mmol) was added into the solution of 4-benzyl 1 - tert-butyl L-aspartate (280 mg, 1 .00 mmol) and triethylamine (0.28 mL, 2.0 mmol) in 5 mL DMF at room temperature, the reaction was stirred overnight, then concentrated and the residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield 400mg, 75%. lon(s) found by LCMS: M+H = 479.3.

Step b. Preparation of methyl (4S,8S,11 S)-8-[2-(benzyloxy)-2-oxoethyl]-4,11-dihexyl-3,6,9-trioxo-2- oxa-5,7,10-triazadodecan-12-oate

4-benzyl 1 -ferf-butyl A/-{[(2S)-1 -methoxy-1 -oxooctan-2-yl]carbamoyl}-L-aspartate (400 mg, 0.83 mmol) was treated with 5mL trifluoroacetic acid in 10mL DCM and stirred for half hour and then concentrated and dried under high vacuum and used for next step without purification.

The above residue was re-dissolved into 20 mL DMF, followed by addition of methyl (2S)-2-amino- octanoate HCI salt (200 mg, 1 mmol), EDC (200 mg, 1 mmol), HOBT (150 mg, 1 mmol), Hunig's base (0.28 mL, 2 mmol) at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. lon(s) found by LCMS: M+H = 578.3.

Step c. Preparation of N-[(2S)-1-methoxy-1-oxooctan-2-yl]-N~2~-{[(2S)-1-methoxy-1-oxooctan-2- yl]carbamoyl}-L-alpha-asparagine

Methyl (4S,8S,1 1 S)-8-[2-(benzyloxy)-2-oxoethyl]-4,1 1 -dihexyl-3,6,9-trioxo-2-oxa-5,7,10- triazadodecan-12-oate (150 mg, 0.26 mmol) was dissolved into 5 mL 5% H20 in MeOH, then 20 mg of 5% Palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for 2hours. The palladium charcoal was removed by filtration, and the filtrate was concentrated and used for next step without purification.

Step d. Preparation of methyl (2S,6S)-2-hexyl-6-{[(2S)-1-methoxy-1-oxooctan-2-yl]carbamoyl}-4,8- dioxo-14-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-12-oxa-3,5,9-triazatetradecan-

1-oate

The residue from Step c. was re-dissolved into 5 mL DMF, followed by addition of INT-1 (65 mg,

0.26 mmol), EDC (1 00 mg, 1 mmol), HOBT (75 mg, 0.5 mmol), Hunig's base (0.14 mL, 1 mmol) at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. lon(s) found by LCMS: M+H = 721 .4.

Step e. Preparation of (2S,6S)-6-{[(1S)-1-carboxyheptyl]carbamoyl}-2-hexyl-4,8-dioxo-14- {[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-12-oxa-3,5,9-triazatetradecan-1-oic acid (non-preferred name)

Methyl (2S,6S)-2-hexyl-6-{[(2S)-1 -methoxy-1 -oxooctan-2-yl]carbamoyl}-4,8-dioxo-14-

{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-12-oxa-3,5,9-triazatetradecan-1 -oate (0.18 mg, 0.25 mmol), in methanol (1 mL) was treated with a solution of lithium hydroxide (0.024 g, 1 .0 mmol), in water (1 mL) and THF (1 mL), then stirred at room temperature for 30 minutes. The reaction was quenched by drops of acetic acid. The desired product was isolated directly by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as modifier. The product was obtained as an oil. lon(s) found by LCMS: M+H = 693.4.

Step f. Preparation of Compound 93

The title compound was prepared analogously using the standard procedure A and B of Example

102. lon(s) found by LCMS: [M/3]+1 =928.2, [M/4]+1 =696.4, [M/5]+1 =557.3, [M/6]+1 =464.6.

Note: The absolute configuration of the trans-cyclopropane- 1 ,2-dicarboxamide is arbitrarily assigned but the product is derived from the more polar trans-cyclopropane-(bis-octanoate tert-butyl ester)

Step a. Synthesis of 1 -benzyl 2,3-di-ferf-butyl 1 -methyl (2S,3S)-cyclopropane-1 ,1 ,2,3- tetracarboxylate

Triphenylarsine (3.17 g, 1 1 mmol) was dissolved in anhydrous EtOH (50 mL) and mixed with benzyl methyl malonate (4.6 g, 22.09 mmol). After the solution was cooled in an ice-water bath, it was added with di-tert-butyl acetylene dicarboxylate (5 g, 22.09 mmol). The reaction mixture was stirred at 0 °C to room temperature for 10 days. EtOH was removed by rotary evaporation, and the residue was purified through silica gel column chromatography (220 g, 0 to 10 % EtOAc and hexane). Yield 9.2 g, 85.9 %. Ion found by LCMS: [M - 2tBu + H]⁺ = 323.

Step b. Selective hydrolysis of tert-butyl ester

The step-a product (9 g, 20.7 mmol) was dissolved in a mixture of DCM (30 mL), TFA (30 mL) and thioanisole (7.7g, 62.2 mmol). After the solution was stirred for 1 day, it was concentrated by rotary evaporation. The residue was purified by RPLC (150 g, 5 to 100 % acetonitrile and water). Yield 6.28 g, 94 %. Ion found by LCMS: [M + H]⁺ = 323. Step c. Synthesis of 1-benzyl 1-methyl (2ff,3/¾-2,3-bis{[(2S)-1-ferf-butoxy-1-oxooctan-2- yl]carbamoyl}cyclopropane-1 ,1-dicarboxylate and its isomer

A mixture of the step-b (646 mg, 2 mmol) and ferf-butyl (2S)-2-aminooctanoate (947 mg, 4.4 mmol) was dissolved in anhydrous DMF (3 m) and DIPEA (1 .14 g, 8.8 mmol). The resulting mixture was drop-wise added with a solution of HATU (1 .67 g, 4.4 mmol) in DMF (5 mL) via syringe pump at a rate of 10 ml/hr. After the addition, an additional amount of DIPEA (600 mg) was added, and the reaction was continued overnight. It was purified by RPLC (1 00 g, 20 to 90 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 374.9 mg of the desired product as more polar isomer (0.523 mmol, 26.1 %). Ions found by LCMS: [M - 2tBu + H]⁺ = 605 and [M + H]⁺ = 717. A less polar isomer was also obtained.

Step d. Selective hydrolysis of benzyl ester

The step-c more polar product was dissolved in EtOAC (~ 20 mL). The solution was added with Pd/C and stirred under hydrogen for 5 hours. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried under high vacuum. Yield 300.6 mg, 91 .2%. Ion found by LCMS: [M - 2tBu + H]⁺ = 515.

Step e. Synthesis of methyl (2S,3¾-2,3-bis{[(2¾-1-fert-butoxy-1-oxooctan-2-yl]carbamoyl}-1-[(2-{2- [(6-deoxy-a-L-mannopyranosyl)oxy]ethoxy}ethyl)carbamoyl]cyclopropane-1-carboxylate

To a solution of a mixture of the step-d product (288 mg, 0.56 mmol) and HATU (254.6 mg, 0.67 mmol) in anhydrous DMF (2 mL) was added with DIPEA (130 mg, 1 mmol). After the resulting mixture was stirred for 10 minutes, it was added with INT-1 (185 mg, 0.35 mmol). The reaction was stirred for 1 hour, then purified by RPLC (40 g, 20 to 75 % acetonitrile and water). Yield 287.4 mg, 59.7 %. Ion found by LCMS: [M + H]⁺ = 860. Step f. Selective hydrolysis of di-tert-butyl ester

The step-e product was dissolved in TFA (~ 2 mL) and thioanisole (0.2 mL). The solution was stirred at room temperature overnight. It was concentrated by rotary evaporator, and the residue was purified by RPLC (40 g, 0 to 40 % acetonitrile and water). Yield 1 1 0.8 mg, 44.4 %. Ion found by LCMS: [M + H]⁺ = 748

Step g. Synthesis of deca-Boc Compound 94 precursor

A mixture of INT-18 (336 mg, 0.2148 mmol) and the step-f product (78.7 mg, 0.1053 mmol) was dissolved in anhydrous DMF (1 mL) and DIEPA (65 mg, 0.5 mmol). After the mixture was cooled in an ice-water bath, it was drop-wise added with a solution of HATU (87.4 mg, 0.23 mmol) in DMF (0.5 mL) via syringe pump at a rate of 0.5 ml/hr. The reaction was then directly purified by RPLC (40 g, 20 to 96 % MeOH and water, using 0.1 TFA as modifier). Yield 338 mg, 83.6 %. Ions found by LCMS: [(M -3Boc + 3H)/3]⁺ = 1 1 81 , [(M - 4Boc + 3H)/3]⁺ = 1 147, [(M - 5Boc + 3H)/3]⁺ = 1 1 14, [(M - 10Boc + 3H)/3]⁺ = 947.

Step h. Removal of the Boc groups Step-g product (338 mg, 0.088 mmol) was dissolved in TFA (~ 2 ml_). After the solution was stirred for 15 minutes, it was concentrated by rotary evaporation and purified by RPLC (50 g, 0 to 30 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 281 .3 mg, 80.4 %. Ions found by LCMS: [(M + 4H)/4]⁺ = 71 0, [(M + 5H)/5]⁺ = 568, [(M + 6H)/6]⁺ = 474, [(M +7H)/7]⁺ = 406.

Step i. Hydrolysis of methyl ester

Step-h product (281 .3 mg, 0.0707 mmol) was dissolved in MeOH (~ 1 mL). After the solution was cooled in ice-water bath, it was added with a solution of LiOH (25 mg, 1 mmol) in water (1 mL). The reaction was stirred for 1 hour and acidified with 4 N HCI solution in dioxane (0.26 mL). It was then directly purified by RPLC (50 g, 0 to 32 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 140 mg, 49.9 %. Ions found by LCMS: [(M + 4H)/4]⁺ = 706, [(M + 5H)/5]⁺ = 565, [(M + 6H)/6]⁺ = 471 , [(M + 7H)/7]⁺ = 404.

Example 180: Synthesis of Compound 95

The compound was prepared by an analogous procedure as that used to prepare Compound 94 except the less polar isomer from Step c was used instead of the more polar diastereomer. Ions found by LCMS: [(M + 4H)/4]+ = 706, [(M + 5H)/5]+ = 565, [(M + 6H)/6]+ = 471

Example 181 : Synthesis of Compound 96

Step a. Preparation of di-tert-butyl 16-{N~2~,N~2~-bis[2-(benzyloxy)-2-oxoethyl]-N,N-bis(17,17- dimethyl-15-oxo-3,6,9,12,16-pentaoxaoctadecan-1-yl)-L-alpha-asparaginyl}-4,7,10,13,19,22,25,28- octaoxa-16-azahentriacontane-1 ,31 -dioate

N,N-bis[2-(benzyloxy)-2-oxoethyl]-L-aspartic acid (150 mg, 0.28 mmol) and di-tert-butyl

4,7,10,13,19,22,25,28-octaoxa-16-azahentriacontane-1 ,31 -dioate (1 80mg, 0.28mmol) and triethylamine (0.1 mL, 0.7 mmol) were dissolved into 5ml_ DMF, then HATU (120 mg, 0.3mmol) in 1 ml_ DMF was added into the resultant solution by syringe pump over 1 hours, the resultant solution was concentrated and isolated directly by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as modifier. The product was obtained as an oil. lon(s) found by LCMS: [M+2H/2]=823.0.

Step b. Preparation of benzyl (25S)-26-[2-(benzyloxy)-2-oxoethyl]-25-[bis(15-oxo-21- {[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-3,6,9,12,19-pentaoxa-16-azahenicosan- 1-yl)carbamoyl]-7,23-dioxo-22-(15-oxo-21-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2- yl]oxy}-3,6,9,12,19-pentaoxa-16-azahenicosan-1-yl)-1-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6- methyloxan-2-yl]oxy}-3,10,13,16,19-pentaoxa-6,22,26-triazaoctacosan-28-oate

Di-tert-butyl 1 6-{N~2~,N~2~-bis[2-(benzyloxy)-2-oxoethyl]-N,N-bis(17,17-dimethyl-15-oxo- 3,6,9,12,16-pentaoxaoctadecan-1 -yl)-L-alpha-asparaginyl}-4,7,10,13,19,22,25,28-octaoxa-16- azahentriacontane-1 ,31 -dioate (500 mg, 0.3 mmol) was treated with 5 mL trifluoroacetic acid in 5 mL DCM and stirred for half hour and then concentrated and dried under high vacuum used for next step without purification. The above residue was re-dissolved into 10 mL DMF, followed by addition of by addition of INT-1 (0.3 g, 1 .2 mmol), Hunig's base (0.3 mL, 2 mmol), then HATU (460 mg, 1 .2 mmol) in 5 mL DMF was added into the resultant solution by syringe pump over 2 hours, the resultant solution was concentrated and isolated directly by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 10% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as modifier. The product was obtained as an oil. lon(s) found by LCMS: [M+2H/2]=1 177.1 , [M+3H/3]=785.1 , [M+4H/4]=589.1 , [M+5H/5]=471 .4.

Step c. Preparation of (25S)-25-[bis(15-oxo-21-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2- yl]oxy}-3,6,9,12,19-pentaoxa-16-azahenicosan-1-yl)carbamoyl]-26-(carboxymethyl)-7,23-dioxo-22- (15-oxo-21-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-3,6,9,12,19-pentaoxa-16- azahenicosan-1-yl)-1-{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-3,10,13,16,19- pentaoxa-6,22,26-triazaoctacosan-28-oic acid

Benzyl (25S)-26-[2-(benzyloxy)-2-oxoethyl]-25-[bis(15-oxo-21 -{[(2R,3R,4R,5R,6S)-3,4,5- trihydroxy-6-methyloxan-2-yl]oxy}-3,6,9,12,1 9-pentaoxa-16-azahenicosan-1 -yl)carbamoyl]-7,23-dioxo-22- (15-0X0-21 -{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-3, 6,9, 12,19-pentaoxa-1 6- azahenicosan-1 -yl)-1 -{[(2R,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy}-3,10,13,1 6,1 9- pentaoxa-6,22,26-triazaoctacosan-28-oate (120 mg, 0.05 mmol) was dissolved into 5 mL 5% H20 in MeOH, then 20 mg of 5% Palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for 2hours. The palladium charcoal was removed by filtration, and the filtrate was concentrated and used for next step without purification.

Step d. Preparation of Compound 96

The title compound was prepared analogously using the standard procedure A and B of Exampli 102. lon(s) found by LCMS: [M+4H/4]=1 136.9, [M+5H/5]=909.7, [M+6H/6]=758.3, [M+7H/7]=650.1 , [M+8H/8]=568.9.

Example 182: Synthesis of Compound 97

Step a. Synthesis of Asn-INT-16

To a solution of INT-16 (464 mg, 0.34 mmol) and Z-L-asparagine (109.2 mg, 0.41 mmol) in anhydrous NMP (2 mL) was added HATU (155.8 mg, 0.41 mmol). The resulting mixture was stirred for 15 minutes, then added with DIPEA (65 mg, 0.5 mmol). After the reaction was stirred for 1 hour, it was diluted with MeOH (~ 15 mL) and treated with Pd/C. The mixture was stirred under hydrogen overnight. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation. The residue was purified by RPLC (50 g, 15 to 80 % MeOH and water). Yield 296.9 mg, 59.1 %. Ion found by LCMS: [(M - Boc + 2H)/2]⁺ = 689. Step b. Synthesis of octa-Boc benzyl bis(2-amino-2-oxoethyl)carbamate Compound 97 precursor

To a solution of step-a product (150 mg, 0.1015 mmol) and (2S,2'S)-2,2'- ({[(benzyloxy)carbonyl]azanediyl}bis[(1 -oxoethane-2,1 -diyl)azanediyl])dioctanoic acid (27.5 mg, 0.05 mmol) in anhydrous DMF (1 mL) was added with DIPEA (65 mg, 0.5 mmol) and drop-wise added with a solution of HATU (20.9 mg, 0.55 mmol) in DMF (0.5 mL) via syringe pump at a rate of 1 ml/hr. The reaction was stirred for 15 more minutes and directly purified by RPLC (50 g, 20 to 100 % MeOH and water). Yield 1 16.6 mg, 67.2 %. Ions found by LCMS: [(M - 3Boc + 3H)/3]⁺ = 1057, [(M - 4Boc + 3H)/3]⁺ =

1023, [(M - 5Boc +3H)/3]⁺ = 990.

Step c. Removed of the Cbz protection

The step-b product was dissolved in MeOH (20 mL) and Pd/C was added. The mixture was stirred under hydrogen overnight. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation to dryness. Yield 104.6 mg, 93.3%. Ions found by LCMS: [(M - 2Boc + 3H)/3]⁺ = 1 045, [(M - 3Boc + 3H)/3]⁺ = 1012, [(M - 4Boc + 3H)/3]⁺ = 978.

Step d. Synthesis of Octa-Boc-Compound 97 precursor

A mixture of step-c product (104.6 mg, 0.0314 mmol) and INT-2 (71 mg, 0.2 mmol) was dissolved in anhydrous DMF (1 mL) and DIPEA (42 mg). After the solution was cooled in an ice-water bath, it was drop-wise added with a solution of HATU (76 mg, 0.2 mmol) in DMF (0.5 mL) via syringe pump at a rate of 1 ml/hr. The reaction was stirred for 15 more minutes and directly purified by RPLC (50 g, 20 to 100 % MeOH and water). Yield 1 15 mg, 96.7 %. Ions found by LCMS: [(M - 3Boc + 3H)/3]⁺ = 1 123, [(M - 6BOC + 3H)/3]⁺ = 1023.

Step e. Removal of the Boc protection

Step-d product (1 1 5 mg, 0.031 mmol) was dissolved in TFA:DCM (1 :1 , - 1 mL). The solution was stirred for 15 minutes, then directly purified by RPLC (50 g, 0 to 35 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 45 mg, 38.4 %. Ions found by LCMS: [(M + 3H)/3]⁺ =956, [(M + 4H)/4]⁺ = 717, [(M + 5H)/5]⁺ = 574, [(M + 6H)/6]⁺ = 478. Example 183: Synthesis of Compound 98

The title compound was prepared analogously to Compound 97. Ions found by LCMS: [(M + 3H)/3]⁺ = 947, [(M + 4H)/4]⁺ = 710, [(M + 5H)/5]⁺ = 568, [(M + 6H)/6]⁺ = 474.

Example 184: Synthesis of Compound 99

The title compound was prepared from INT-44 and INT-54 in a similar manner as described in Example 37. Yield: 52%. LC/MS [M+4H/4]+ = 884.8. xample 185: Synthesis of Compound 100

The title compound was prepared analogously to Compound 97. Ions found by LCMS: [(M + 3H)/3]⁺ = 956.7, [(M + 4H)/4]⁺ = 718, [(M + 5H)/5]⁺ = 574.

Example 186: Synthesis of Compound 101

INT-59 (0.289 g, 0.0985 mmol), INT-18 (0.323 g, 0.207 mmol) and DIPEA (0.051 mL, 0.30 mmol) were mixed in 5 mL of DMF. HATU (0.0936 g, 0.246 mmol) was added via syringe pump (1 mL, 1 ml/hr). The reaction solution was stirred for another hour after addition. DMF was removed and the residue was purified by prep-HPLC to give per-Boc-Compound 91 , 0.281 g (47%). Positive mass ions were found as (M - 4Boc + 8H⁺)/8: 703.6, (M - 4C₆Hii0₄ + 5H⁺)/5: 1087.0, tr = 2.09 minutes with 8 min (40-95%) method. The Boc groups of Per-Boc-Compound 1 01 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Preparative HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M - 4C₆Hii0₄ + 5H⁺)/5: 888.3, (M -3C₆Hii0₄ + 5H⁺)/5: 917.7, (M - 2C₆Hii0₄ + 5H⁺)/5: 947.2, (M - C₆Hii0₄ + 5H⁺)/5: 976.8, (M + 5H⁺)/5: 1005.7, (M + 4H⁺)/4: 1256.7] at tr = 2.26 min with 8 min (5-95%) method for Compoud 101 .

Example 187: Synthesis of Compound 102

The title compound was prepared analogously to compound Compound 97. Ions found by LCMS: [(M + 4H)/4]⁺ = 733.9, [(M + 5H)/5]⁺ = 587, [(M + 6H)/6]⁺ = 489, [(M + 7H)/7]⁺ = 420.

Step a. Preparation of methyl (2S)-2-({A/⁶-[(benzyloxy)carbonyl]-A -(fert-butoxycarbonyl)-L- lysyl }am i no)octanoate

Methyl (2S)-2-amino-octanoate (350 mg, 2 mmol), ferf-butyl /^-[(benzyloxyJcarbonylJ-L-lysinate

(770 mg, 2 mmol), EDC (500 mg, 2.5 mmol), HOBT (450 mg, 3 mmol), Hunig's base (0.7 mL, 5 mmol) were mixed in 10 mL DMF at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 1 00% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product 460 mg (75%) was obtained as an oil. lon(s) found by LCMS: M+H = 536.3. Step b. Preparation of methyl (2¾-2-{[(2S)-6-{[(benzyloxy)carbonyl]amino}-2-({[(2S)-1-methoxy-1- oxooctan-2-yl]carbamoyl}amino)hexanoyl]amino}octanoate

Methyl (2S)-2-({/V⁶-[(benzyloxy)carbonyl]-/v²-(ferf-butoxycarbonyl)-L-lysyl}amino)octanoate (540mg, 1 .0 mmol) was treated with 2 mL trifluoroacetic acid in 2mL DCM and stirred for half hour and then concentrated and dried under high vacuum used for next step without purification.

The deprotected products from the above step were dissolved into 5 mL DMF and methyl (2S)-2- isocyanatooctanoate (200 mg, 1 mmol) and triethylamine (0.7 mL, 5 mmol) were added at room temperature. The reaction was stirred overnight then concentrated and the residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. Yield 460 mg, 75%. lon(s) found by LCMS: M+H = 635.4.

Step c. Preparation of methyl (2S)-2-({[(2S)-6-amino-1-{[(2S)-1-methoxy-1-oxooctan-2-yl]amino}-1- oxohexan-2-yl]carbamoyl}amino)octanoate

Methyl (2S)-2-{[(2S)-6-{[(benzyloxy)carbonyl]amino}-2-({[(2S)-1 -methoxy-1 -oxooctan-2- yl]carbamoyl}amino)hexanoyl]amino}octanoate (150 mg, 0.24 mmol) was dissolved into 5 mL 5% H20 in MeOH, then 50 mg of 5% Palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere for 2hours. The palladium charcoal was removed by filtration. The residue was used for next step without purification. Step d. Preparation of methyl (2S)-2-{[(2S)-6-{4-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-4-oxobutanamido}-2-({[(2S)-1 -methoxy-1 -oxooctan-2- yl]carbamoyl}amino)hexanoyl]amino}octanoate

The methyl (2S)-2-({[(2S)-6-amino-1 -{[(2S)-1 -methoxy-1 -oxooctan-2-yl]amino}-1 -oxohexan-2- yl]carbamoyl}amino)octanoate from previous step was redissoved into 5mL DMF, followed by addition of INT-2 (83 mg, 0.24 mmol), EDC (100 mg, 0.5 mmol), HOBT (80 mg, 0.5 mmol), Hunig's base (0.14 mL, 1 mmol) at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. lon(s) found by LCMS: M+H = 834.5.

Step e. Preparation of (2S)-2-[({(16S,19S)-19-carboxy-1-[(6-deoxy-alpha-L-mannopyranosyl)oxy]- 7,10,17-trioxo-3-oxa-6,11 ,18-triazapentacosan-16-yl}carbamoyl)amino]octanoic acid

Methyl (2S)-2-{[(2S)-6-{4-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-4- oxobutanamido}-2-({[(2S)-1 -methoxy-1 -oxooctan-2-yl]carbamoyl}amino)hexanoyl]-amino}octanoate (0.050 g, 0.06 mmol) in methanol (1 mL) was treated with a solution of lithium hydroxide (0.012 g, 0.5 mmol), in water (1 mL) and THF (1 mL), then stirred at room temperature for 30 minutes. The reaction was quenched by drops of acetic acid. The desired product was isolated directly by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% acetonitrile and water, using 0.1 % Trifluoacetic acid as modifier. Yield of oil product 48mg, 100% yields. lon(s) found by LCMS: M+H = 806.5.

Step f. Preparation of Compound 103

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3]=965.9 [M+4H/4]=724.7, [M+5H/5]=579.9, [M+6H/6]=483.5.

Step a. Synthesis of 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}azanediyl)diacetic acid

Lithium hydroxide (480 mg, 20 mmol) in 10mL H2O was added to the solution of diethyl 2,2'-({2- [(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}azanediyl)-diacetate (4.80 g, 10 mmol) in mix solvent of 10 mL H2O, 20 mL MeOH and 20 mL THF. The resultant solution was stirred for 1 hour at room temperature. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoro-acetic acid as the modifier. The product was obtained as an oil. LC/MS 425.2 [M+H]⁺ Step b. Synthesis of dibenzyl 14-{2-[(2-{2-[(6-deoxy-alpha-L- mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}-4,12,16,24-tetraoxo-8,20-dioxa- 5,11 ,14,17,23-pentaazaheptacosane-1 ,27-dioate

To the solution 2,2'-({2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}azanediyl)diacetic acid (1 .3 g, 3 mmol) and 4-(benzyloxy)-4-oxobutanoic acid (1 .8 g, 6.1 mmol) in 30 mL DMF was added EDC (1 .4 g, 7 mmol), HOBT (1 .0 g, 7 mmol), Hunig's base (1 .4 mL, 10 mmol) at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. LC/MS 977.5 [M+H]⁺

Step c. Synthesis of 14-{2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2- oxoethyl}-4,12,16,24-tetraoxo-8,20-dioxa-5,11 ,14,17,23-pentaazaheptacosane-1 ,27-dioic acid

Dibenzyl 14-{2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}- 4,12, 16,24-tetraoxo-8,20-dioxa-5, 1 1 ,14,17,23-pentaazaheptacosane-1 ,27-dioate (816 mg, 0.836 mmol) was dissolved into 5 mL MeOH and 5 mL ethyl acetate, then 0.5 g of 5% Palladium on charcoal was added above solution, and the mixture was stirred at room temperature under the hydrogen atmosphere overnight. The palladium charcoal was removed by filtration after completed the reaction by LCMS. The filtrate was concentrated and used next step without any purification. LC/MS, 797.4 [M+H]+ .

Step d. Synthesis of Compound 104

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3]=1056.9, [M+4H/4] =792.9, [M+5H/5]=634.6, [M+6H/6]=529.0, [M+7H/7]=453.5.

Example 190: Synthesis of Compound 105

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3]= 1020.2, [M+4H/4] =776.2, [M+5H/5]=613.1 , [M+6H/6]=51 1 .1 .

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3]=924.2, [M+4H/4]=693.4, [M+5H]+1 =554.9, [M+6H/6]=462.6.

The title compound was prepared from INT-38a and INT-65 in a similar manner as described for the synthesis of Compound 64. Yield: 66 %. LC/MS [M+3H/3]+ = 861 .4.

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3]=1023.3, [M+4H/4]=768.4, [M+5H/5]=614.9, [M+6H/6]=512.6.

Step a. Synthesis of D-Ser-INT-11

Z-D-Ser-OH (435.3 mg, 1 .82 mmol) and INT-1 1 (1 .6 g, 1 .51 mmol) were dissolved in anhydrous DMF (2 mL). It was cooled in an ice-water bath and mixed with DIPEA (260 mg) followed by dropwise addition of a solution of HATU (722 mg, 1 .9 mmol) in DMF (2 mL) via syringe pump at a rate of 4 ml/hr. After the addition, the reaction was directly purified by RPLC (100 g, 30 to 85 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (Ion found by LCMS: [(M - 2Boc + 2H)/2]⁺ = 542). The material was re-dissolved in MeOH (~ 30 ml_) and added with Pd/C. The mixture was stirred under hydrogen overnight. The Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried under high vacuum. Yield 1 .23 g, 70.7 %. Ion found by LCMS: [(M -Boc + 2H)/2]⁺ = 525.

Step b. Synthesis of Thr-(D)-Ser-INT-11

Z-thr-OH (329.3 mg, 1 .3 mmol) and step-a product (1 .23 g, 1 .07 mmol) were dissolved in anhydrous DMF (2 ml_). After the solution was cooled in an ice-water bath, it was mixed with HATU (418 mg, 1 .1 mmol). The mixture was stirred for 10 minutes, then added with DIPEA (195 mg, 1 .5 mmol). The reaction was stirred for 1 hour and directly purified by RPLC (100 g, 30 to 85 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (Ion found by LCMS: [(M - 2Boc + 2H)/2]⁺ = 593). The material was re-dissolved in MeOH (~ 30 mL) and added with Pd/C. The mixture was stirred under hydrogen overnight. The Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried under high vacuum. Yield 1 .03 g, 77 %. Ion found by LCMS: [(M - Boc + 2H)/2]⁺ = 576.

Step c. Synthesis of Dab-Thr-(D)-Ser-INT-11

Z-thr-OH (123.5 mg, 0.487 mmol) and step-b product (508 mg g, 0.406 mmol) were dissolved in anhydrous DMF (1 mL) and mixed with HATU (190 mg, 0.5 mmol). After the mixture was stirred for 10 minutes, DIPEA (195 mg, 1 .5 mmol) was added. The reaction was stirred for 1 hour and directly purified by RPLC (100 g, 30 to 85 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (Ion found by LCMS: [(M - 3Boc + 2H)/2]⁺ = 593). The material was re- dissolved in MeOH (~ 30 mL) and added with Pd/C. The mixture was stirred under hydrogen overnight. The Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation and further dried under high vacuum. Yield 448.7 mg, 81 .8 %. Ion found by LCMS: [(M - 2Boc + 2H)/2]⁺ = 576.

Step d. Synthesis of hexa-Boc-Compound 109 precursor

To a solution of step-c product (273.2 mg, 0.2 mmol) and INT-43 (46 mg, 0.096 mmol) in anhydrous DMF (1 mL) and DIPEA (65 mg, 0.5 mmol) was drop-wise added a solution of HATU (76 mg, 0.2 mmol) in DMF (0.5 mL) via syringe pump at a rate of 1 ml/hr. The reaction was stirred for 15 more minutes and directly purified by RPLC (50 g, 20 to 100 % MeOH and water). Yield 248 mg, 75.6 %. Ions found by LCMS: [(M - Boc + 3H)/3]+ = 1046.7, [(M - 3Boc + 3H)/3]⁺ = 1 038, [(M - 5Boc + 3H)/3]⁺ = 971 .

Step e. Removal of the Boc protection

Step-d product (248 mg, 0.072 mmol) was dissolved in TFA (~1 mL). The solution was stirred for

15 minutes, then directly purified through HPLC (5 to 26 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 71 .4 mg, 29.4 %. Ions found by LCMS: [(M + 4H)/4]⁺ =704, [(M + 5H)/5]⁺ = 563.7. Example 195: Synthesis of Compound 110

Step a. Synthesis of Methyl (2S)-2-Aminooctanoate

(2S)-2-Aminooctanoic acid (3.0 g, 18.84 mmol) was stirred and heated in methanol (40 mL) and sulfuric acid (2.0 mL) at 75 °C in a sealed tube overnight. Next day, the reaction mixture was cooled down to room temperature then with vigorously stirring the reaction was quenched with saturated NaHCCb aqueous solution (50 mL). Upon the gas evolution ceased and pH 6, methanol was removed under reduced pressure. The aqueous layer was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with brine, dried over solid Na2S04 and filtered then concentrated. The residue was used in the next step without purification. Ion found by LC/MS [M+H]+ = 174.2.

Step b. Synthesis of pyrrolidine intermediate (a) more polar isomer and pyrrolidine intermediate (b) less polar isomer

To a solution of racemic-trans1 -(tert-butoxycarbonyl)pyrrolidine-3,4-dicarboxylic acid (0.3 g, 1 .16 mmol), methyl (2S)-2-aminooctanoate (0.44 g, 2.55 mmol), and diisopropylethylamine (896.6 mg, 6.94 mmol) in DMF (5.7 mL) was added a solution of HATU (967.95 mg, 2.55 mmol) in DMF (4 mL) via a syringe pump at a rate of 2.5 mL/hr. After the addition, the reaction mixture was stirred for an additional of 1 h at room temperature. The reaction mixture was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 50% acetonitrile/water, using 0.1 % TFA as modifier to yield two diastereomers: the more polar diastereomer (a) (381 .8 mg, 58%) and the less polar diastereomer (b) (344.3 mg, 52%). Ion found by LC/MS [M-Boc+H]+ = 470.4 (loss of 1 boc group) for both Boc-protected compounds, methyl 2-[((3R,4R)-4-{N[(1 S)-1 - (methoxycarbonyl)heptyl]carbamoyl}-1 -[(tert-butyl)oxycarbonyl]pyrrolidine-3- yl)carbonylamino](2S)ocatanoate. Note: the isomers were taken forward separately in subsequent syntheses. Step c. Removal of the Boc Group

The Step-b product (more polar isomer) (381 .8 mg, 0.67 mmol) was stirred in TFAiChteC (1 :1 , 12 ml_) at room temperature then volatiles were removed under reduced pressure. The residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 5 to 100% methanol and Dl water to yield the title compound (201 .0 mg, 64%). Ion found by LC/MS [M+H]+ = 470.4.

Step d. Acylation with I NT- 2

A solution of HATU (198.21 mg, 0.52 mmol) in DMF (0.8 mL) was added via a syringe pump at a rate of 2.5 mL/hr into a solution of the Step-c Product (204 mg, 0.43 mmol), INT-2 (183.15 mg, 0.52 mmol), diisopropylethylamine (168.29 mg, 1 .30 mmol) in DMF (4 mL). After the addition, the reaction mixture was stirred for an additional hour then purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 80% acetonitrile and water, using 0.1 % TFA as modifier to yield the title compound (221 .5 mg, 64%). Ion found by LC/MS [M+H]+ = 803.4. Step e. Hydrolysis of the Methyl Esters

The Step-d Product (222.0 mg, 0.28 mmol) and LiOH (17.8 mg, 0.74 mmol) were stirred in MeOH:THF:H20 (1 :1 :2, 16 mL) at room temperature for 3 hours. After the reaction was complete, it was quenched with acetic acid to pH 4 then it was concentrated. The residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0 to 45% acetonitrile and water, using 0.1 % TFA as modifier) to yield the title compound (161 mg, 75%). Ion found by LC/MS [M-H]+ = 773.3.

Step f. Synthesis of Compound 110

A solution of HATU (53.98 mg, 0.14 mmol) in DMF (0.4mL) was added via a syringe pump at a rate of 2.5 mL/hr into a solution of the Step-e Product (50.0 mg, 0.065 mmol), INT-18 (221 .99 mg, 0.14 mmol) and diisopropylethylamine (50 mg, 0.39 mmol) in DMF (1 .3 mL). The reaction mixture was stirred for an additional hour then purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0 to 30 to 00% methanol and water to yield the Boc- protected intermediate. The Boc-protected intermediate was stirred in TFA:DCM (1 :1 , 6 mL) at room temperature for 20 minutes. After the reaction was completed, the reaction mixture was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid

chromatograph eluted with 5 to 45% acetonitrile and water, using 0.1 % TFA as modifier to yield the title compound (1 85.7 mg, 72%). lon(s) found by LC/MS [(M+4H)/4]+ = 71 6.8, [(M+5H)/5]+ = 573.73 and [(M+6H)/6]+ = 478.28.

Example 196: Synthesis of Compound 111

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3] = 1033.6, [M+4H/4] =776.2, [M+5H/5]=620.2, [M+6H/6]=517.8.

The title compound was prepared analogously to Example 37, where Cbz-L-aminodecanoic acid was used in place of Cbz-L-amino-octanoic acid for the first step of that sequence, lon(s) found by LCMS: (M+3H)/3 = 965.3, (M+4H)/4 = 724.3, (M+5H)/5 = 579.7

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3] = 1020.2, [M+4H/4] =776.2, [M+5H/5]=613.1 , [M+6H/6]=51 1 .1 .

Example 199: Synthesis of Compound 114

A solution of HATU (50.06 mg, 0.13 mmol) in DMF (0.4 mL) was added via a syringe pump at a rate of 2.5 mL/hr into a solution of INT-64 (191 .0 mg, 0.13 mmol), INT-66 (48.30 mg, 0.062 mmol) and diisopropylethylamine (48.58 mg, 0.38 mmol) in DMF (1 mL). After the addition, the reaction was stirred for an additional hour at room temperature then purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0 to 30 to100% methanol and water, using 0.1 % TFA as modifier to yield the Boc-protected intermediate. The Boc-intermediate was stirred in TFA:CH2Cl2 (1 :1 , 6 mL) at room temperature for 30 minutes then the volatiles were removed under reduced pressure. The residue was purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5 to 45% acetonitrile and water, using 0.1 % TFA as modifier to yield the title compound as TFA salt (89.4 mg, 38%). lon(s) found by LC/MS [(M+4H)/4]+ = 710.6, [(M+5H)/5]+ = 568.8, [(M+6H)/6]+ = 474.2.

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3] = 1006.3, [M+4H/4] =754.7, [M+5H/5]=604.8, [M+6H/6]=504.1 .

Step a. Synthesis of (2¾-2-{[(benzyloxy)carbonyl]amino}-4-(ureidoureido)butanoic acid

A suspension of (S)-4-amino-2-(benzyloxycarbonylamino)butyric acid (1 .01 g, 4 mmol) in water (4 mL) was heated to dissolved. After cooled to room temperature, the solution was added with potassium cyanate (648.8 mg, 8 mmol). The reaction was stirred overnight, then directly purified by RPLC (1 00 g, 0 to 30 % acetonitrile and water). Yield 966 mg, 81 .8 %. Ion found by LCMS: [M + H]⁺ = 296. Step b. Synthesis of 4-(ureido)butanamide-INT-11

To a solution of INT-1 1 (618 mg, 0.6 mmol) and the step-a product (212.4 mg, 0.72 mmol) in anhydrous DMF (1 mL) and DIPEA (1 10 mg, 0.85 mmol) was drop-wise added a solution of HATU (273.6 mg, 0.72 mmol) in DMF (1 mL) via syringe pump at a rate of 2 ml/hr. After the addition, the mixture was purified by RPLC (100 g, 1 5 to 85 % MeOH and water). The collected fractions were concentrated by rotary evaporation to a white solid (Ions found by LCMS: [(M - 2 Boc + 2H)/2]⁺ = 570, [(M - 3 Boc + 2H)/2]⁺ = 520). The material was dissolved in MeOH (~ 25 mL) and added with Pd/C. The mixture was stirred under hydrogen overnight. Pd/C was filtered off, and the filtrate was concentrated by rotary evaporation to dryness. Yield 520 mg, 71 .9 %. Ions found by LCMS: [(M - Boc + 2H)/2]⁺ = 553. Step c. Synthesis of octa-Boc-Compound 116 precursor

To a solution of the step-b product (240.5 mg, 0.2 mmol) and INT-40 (124.3 mg, 0.092 mmol) in anhydrous DMF (1 mL) and DIPEA (65 mg, 0.5 mmol) was drop-wise added a solution of HATU (76 mg, 0.2 mmol) in DMF (1 mL) via syringe pump at a rate of 2 ml/hr. After the addition, the mixture was purified by RPLC (100 g, 20 to 95 % MeOH and water). Yield 206.8 mg, 60.3%. Ions found by LCMS: [(M - 2Boc + 3H)/3]⁺ = 1 175.7, [(M - 3Boc + 3H)/3]⁺ = 1 142, [(M - 4Boc + 3H)/3]⁺ = 1 109, [(M - 5Boc + 3H)/3]⁺ = 1075, [(M - 6Boc + 3H)/3]⁺ = 1042.

Step d. Removal of the Boc group

Step-c product (206.8 mg, 0.056 mmol) was dissolved in TFA:DCM (1 :1 , ~ 1 .5 mL). The solution was stirred for 15 minutes and purified through HPLC (5 to 25 % acetonitrile and water, using 0.1 % TFA as modifier). Yield 62.8 mg, 29.5 %. Ions found by LCMS: [(M + 3H)/3]⁺ = 975.3, [(M + 4H)/4]⁺ = 731 .9, [(M + 5H)/5]⁺ = 585.7, [(M + 6H)/6]⁺ = 488.3.

Example 202: Synthesis of Compound 117

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3]=915.5, [M+4H/4]=686.9, [M+5H/5]=572.6, [M+6H/6]=477.3.

Example 204: Synthesis of Compound 119

The title compound was prepared in an analogous fashion to the procedure used to prepare Compound 120. lon(s) found by LCMS: [M+3H/3]=1043.3, [M+4H/4]=783.5, [M+5H/5]=627.0,

[M+6H/6]=522.6, [M+7H/7]=448.1 .

Step a. Preparation of benzyl 12-[2-(benzyloxy)-2-oxoethyl]-1-[(6-deoxy-alpha-L- mannopyranosyl)oxy]-9-{2-[(2-{2-[(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)-amino]-2- oxoethyl}-7-oxo-3-oxa-6,9,12-triazatetradecan-14-oate

2,2'-[(2-{bis[2-(benzyloxy)-2-oxoethyl]amino}ethyl)azanediyl]diacetic acid (200 mg, 0.4 mmol) was dissoved into 10 mL DMF, followed by addition of INT-1 (250 mg, 1 mmol), EDC (200 mg, 1 mmol), HOBT (150 mg, 1 mmol), Hunig's base (0.28 mL, 2 mmol) at room temperature. The solution was stirred overnight. The resultant solution was concentrated and purified by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 5% to 100% acetonitrile and water, using 0.1 % trifluoroacetic acid as the modifier. The product was obtained as an oil. lon(s) found by LCMS: M+H = 939.4. Step b. Preparation of 12-(carboxymethyl)-1-[(6-deoxy-alpha-L-mannopyranosyl)oxy]-9-{2-[(2-{2- [(6-deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)amino]-2-oxoethyl}-7-oxo-3-oxa-6,9,12- triazatetradecan-14-oic acid

Benzyl 12-[2-(benzyloxy)-2-oxoethyl]-1 -[(6-deoxy-alpha-L-mannopyranosyl)oxy]-9-{2-[(2-{2-[(6- deoxy-alpha-L-mannopyranosyl)oxy]ethoxy}ethyl)-amino]-2-oxoethyl}-7-oxo-3-oxa-6,9,12- triazatetradecan-14-oate (200 mg, 0.02 mmol) was dissolved into 10 mL 5% H20 in MeOH, then 100 mg of 5% Palladium on charcoal was added, and the mixture was stirred at room temperature under a hydrogen atmosphere for 2 hours. The palladium charcoal was removed by filtration, and the filtrate was concentrated and used for next setp without purification, lon(s) found by LCMS: M+H=759.3.

Step c. Preparation of Compound 120

The title compound was prepared analogously using the standard procedure A and B of Example 102. lon(s) found by LCMS: [M+3H/3]=1035.6, [M+4H/4]=776.9, [M+5H/5]=621 .7, [M+6H/6]=517.3, [M+7H/7]=444.4.

The title compound was prepared analogously using the standard procedure A and B of Exampli 102. lon(s) found by LCMS: [M+3H/3]=1048.3, [M+4H/4] =786.4, [M+5H/5]=629.4, [M+6H/6]=524.6, [M+7H/7]=449.8.

Example 207: Synthesis of Compound 121

Step a. Synthesis of methyl 3-(aminomethyl)-5-bromobenzoate

In a sealed tube, a stirring mixture of methyl 3-bromo-5-(bromomethyl)benzoate (3.08 g, 10.00 mmol) and sodium azide (0.715 g, 1 1 .00 mmol) in DMF (10 mL) was heated at 80 °C for 1 h, when HPLC analysis showed full conversion of the bromide. All the volatiles were evaporated per vacuum techniques. The residue was taken up in DCM (40 mL) and water (40 mL). The water layer was washed with additional DCM (2 x 30 mL). The combined organic layers were dried with brine and solid sodium sulfate. Upon filtration, all the volatiles were evaporated per vacuum techniques, affording 2.73 g of the crude azide derivative in high purity, as confirmed per ¹ H-NMR and LC-MS analysis. ¹ H NMR (DMSO-cfe) δ: 8.03 (t, J = 1 .6 Hz, 1 H), 7.98 - 7.93 (m, 1 H), 7.90 (t, J = 1 .5 Hz, 1 H), 4.60 (s, 2H), 3.88 (s, 3H). This azide (2.73 g) was dissolved under stirring in THF (30 mL) and water (2 mL), and triphenylphosphine (3.148 g, 12.00 mmol) was added. The reaction was stirred at room temperature overnight, allowing gas evolution through a bubbler. All the volatiles were evaporated per vacuum techniques. The desired product was isolated by normal phase liquid chromatography using an Isco CombiFlash liquid chromatograph eluted with 1 % to 20% hexanes and ethylacetate. From the desired fractions, the desired compound was obtained still contaminated with triphenylphenylphosphine oxide. This material was therefore dissolved in DCM (50 mL) and extracted an acidic solution (12 mL of 1 .0 M H2SO4 and 30 mL of water). The water layer was added dropwise to a vigorously stirring solution of saturated sodium bicarbonate (60 mL) and stirring was continued overnight. The obtained mixture was extracted with DCM (3 x 30 mL). The combined organic layers were dried with brine and solid sodium sulfate. Upon filtration, all the volatiles were evaporated per vacuum techniques, affording 1 .31 g, 54%. Ions found by LCMS: (M+H)+ = 244.0; 246.0. Step b. Synthesis of methyl 3-((N-(1 '-(3'-carboxymethyl-5-bromo)phenylene)methylene)methyl)-5- bromobenzoate

A solution of methyl 3-(aminomethyl)-5-bromobenzoate (1 .306 g, 5.350 mmol) and methyl 3- bromo-5-formylbenzoate (1 .238 g, 5.096 mmol) in DCM (10 mL) was evaporated per vacuum techniques and left over high vacuum overnight. The residue was taken up in THF (30 mL) and treated under vigorous stirring with sodium triacetoxyborohydride (3.240 g, 15.29 mmol). This mixture was quenched after 18 h by the addition of a saturated aqueous solution of ammonium chloride (50 mL). The obtained mixture was extracted with DCM (3 x 30 mL). The combined organic layers were dried with brine and solid sodium sulfate. Upon filtration, all the volatiles were evaporated. The desired product was isolated by normal phase liquid chromatography using an Isco CombiFlash liquid chromatograph eluted with 1 % to 50% hexanes and ethylacetate. Yield 1 .61 1 g, 67%. Ions found by LCMS: (M+H)+ = 470.0, 472.1 , 474.0.

Step c. Synthesis of N-Cbz protected methyl 3-((N-(1 '-(3'-carboxymethyl-5- bromo)phenylene)methylene)methyl)-5-bromobenzoate

To a 0^°C stirring solution of methyl 3-((N-(1 '-(3'-carboxymethyl-5- bromo)phenylene)methylene)methyl)-5-bromobenzoate (1 .61 1 g, 3.419 mmol) and DIPEA (0.953 mL, 5.471 mmol) in THF (20 mL) it was added Cbz-CI (0.732 mL, 5.129 mmol) dropwise. The temperature was raised to ambient after 10 minutes and stirring was continued until complete disappearance of the starting amine by LC-MS analysis. All the volatiles were removed per rotatory evaporation. The desired product was isolated by normal phase liquid chromatography using an Isco CombiFlash liquid chromatograph eluted with 1 % to 20% hexanes and ethylacetate. Yield 2.070 g, quant. Ions found by LCMS: (M+H)+ = 603.0, 605.0, 607.0. Step d. Synthesis of N-Cbz protected 3-((N-(1 '-(3'-carboxy-5-phenyl)phenylene)methylene)methyl)- 5-phenylbenzoic acid

Under nitrogen, in a capped microwave reaction vessel stirring mixture of N-Cbz protected methyl 3-((N-(1 '-(3'-carboxymethyl-5-bromo)phenylene)methylene)methyl)-5-bromobenzoate (0.299 g, 0.494 mmol), potassium phenyltrifluoroborate (0.200 g, 1 .087 mmol), sodium carbonate (0.209 g, 1 .976 mmol) and bis(triphenylphosphine)palladium (II) dichloride (0.035 g, 0.049 mmol) in MeCN (4 mL) and water (4 mL) were irradiated with microwaves for 10 mim at 130 °C. Upon cooling, the reaction was treated under stirring with SiliaMetS® (0.200 g, 0.240 mmol) for 1 h, and TFA was added (0.380 mL, 5.0 mmol). Upon filtration, all the volatiles were evaporated per vacuum techniques. The desired product was isolated as a dodeca-TFA salt by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% methanol and water, using TFA as the modifier. Yield 0.072 g, 25% yield. ¹ H NMR (0.500 mL Chloroform-d + 0.050 mL TFA) δ: 8.1 8 (d, J = 5.5 Hz, 2H), 7.82 (d, J = 16.6 Hz, 2H), 7.60 (d, J = 33.5 Hz, 2H), 7.52 - 7.27 (m, 15H), 5.32 (s, 2H), 4.70 (d, J = 19.9 Hz, 4H).

Step e. Synthesis of N-Cbz-deca-Boc-(Compound 121)-EM

A stirring solution of INT-18 (0.429 g, 0.241 mmol), N-Cbz protected 3-((N-(1 '-(3'-carboxy-5- phenyl)phenylene)methylene)methyl)-5-phenylbenzoic acid (0.069 g, 0.121 mmol), and DIPEA (0.128 mL, 0.736 mmol) in DMF (10 mL), was treated with a solution of HATU (0.094 g, 0.247 mmol in 3.0 mL of DMF), dropwise over 30 minutes. After 1 .5 hour, all the volatiles were evaporated per vacuum techniques. The desired product was isolated by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 50% to 100% methanol and water, using no modifier. Yield 0.1 58 g, 37% yield. Ion found by LCMS: [(M-3Boc)+3H]/4 = 841 .2. Step f. Synthesis of deca-Boc-(Compound 121)-EM

N-Cbz-deca-Boc-(Compound 121 )-EM (0.1 58 g, 0.0431 mmol) was dissolved methanol (10 ml_) and suspended with SiliaCat® Pd(0) (0.043 g; 0.009 mmol) under hydrogen atmosphere (~1 atm). When all the starting material was consumed by HPLC analysis, the mixture was filtered over a short Celite pad, and all the volatiles were evaporated per vacuum techniques. HPLC analysis revealed the desired compound in high purity, and this material was used without further purification. Yield 0.1 51 g, 99% yield. Ion found by LCMS: [(M-3Boc)+3H]/4 = 808.8.

Step g. Synthesis of deca-Boc-Compound 121

Prepared in a similar fashion to "Step e. Synthesis of N-Cbz-deca-Boc-Compound 121 -EM" from deca-Boc-Compound 121 -EM (0.151 g, 0.043 mmol), INT-9 (0.045 g, 0.128 mmol), 2,4,6-trimethylpyridine (0.034 mL, 0.261 mmol) in DMF (5 mL) by dropwise addition of HATU (0.050 g, 0.130 mmol in 1 .5 mL of DMF). Yield 0.062 g, 38% yield. Diagnostic peaks from ¹ H NMR (Methanol-ck) δ: 4.71 (br s, 1 H), 2.64 (br s, 2H). Step h. Synthesis of Compound 121

A solution of deca-Boc-Compound 121 (0.062 g, 0.016 mmol), dissolved in DCM (4 mL) and triethylsilane (0.250 mL), was treated with TFA (2 mL), while stirring at room temperature. After 30 minutes, all the volatiles were evaporated per vacuum techniques. The desired product was isolated as the deca-TFA salt by reversed phase liquid chromatography (RPLC) using an Isco CombiFlash liquid chromatograph eluted with 0% to 100% methanol and water, using TFA as the modifier. Yield 0.042 g, 65% yield. Main ions found by HRMS: (M+5H)/5 = 573.0.

Example 208: Synthesis of Compound 122

INT-60 (0.0500 g, 0.1 07 mmol), INT-20 (0.366 g, 0.214 mmol) and DIPEA (0.047 ml_, 0.27 mmol) were mixed in 5 mL of DMF. HATU (0.102 g, 0.269 mmol) was added via syringe pump (1 ml_, 1 ml/hr). The reaction solution was stirred for another hour after addition. DMF was removed and the residue was purified by prep-HPLC to give per-Boc-Compound 122, 0.223 g, 54%. Positive mass ions were found as (M - 4Boc + 8H⁺)/8: 703.6, (M - CeH C - 6Boc + 4H⁺)/4: 772.4, tr = 3.21 minutes with 7 min (60-95%) method. The Boc groups of Per-Boc-Compound 122 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Preparative HPLC with ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M + 5H⁺)/5: 568.3, (M + 4H⁺)/4: 710.1 ] at tr = 1 .01 min with 5 min (5-95%) method for Compound 122.

Example 209: Synthesis of Compound 123

INT-61 (0.0500 g, 0.1 04 mmol), INT-20 (0.3556 g, 0.2086 mmol) and DIPEA (0.045 mL, 0.26 mmol) were mixed in 5 mL of DMF. HATU (0.0991 g, 0.261 mmol) was added via syringe pump (1 mL, 1 ml/hr). The reaction solution was stirred for another hour after addition. DMF was removed and the residue was purified by prep-HPLC to give per-Boc-Compound 123, 0.208 g, 51 %. Positive mass ions were found as (M - CeH C - 3Boc + 4H⁺)/4: 851 .6, (M - CeH C - Boc + 4H⁺)/4: 901 .0, (M - 2Boc + 3H⁺)/3: 1216.8, tr = 2.44 minutes with 5 min (40-95%) method. The Boc groups of Per-Boc-Compound 123 were removed by treating with TFA for less than 1 hour. Most of the TFA was removed and the residue was dissolved in a minimum amount of NMP and loaded onto a Preparative HPLC with

ACN/water (0.1 % TFA). The desired product was collected and lyophilized to give the product as a white powder of TFA salt. Positive mass ions were found as [(M + 6H⁺)/6: 475.0, (M + 5H⁺)/5: 571 .0, (M + 4H⁺)/4: 713.5] at tr = 0.6 min with 5 min (5-95%) method for Compound 123.

Other Embodiments

While the disclosure has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the disclosure following, in general, the principles of the disclosure and including such departures from the present disclosure come within known or customary practice within the art to which the disclosure pertains and may be applied to the essential features hereinbefore set forth. All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publijyacation, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

What is claimed is:

Claims

1 . A compound described by formula (I):

(I)

wherein M1 comprises a first cyclic heptapeptide comprising a linking nitrogen and M2 comprises a second cyclic heptapeptide comprising a linking nitrogen ;

each E is, independently, a monosaccharide or oligosaccharide moiety;

L' is a linker covalently attached to E and to the linking nitrogen in each of M1 and M2;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof. 2. The compound of claim 1 , wherein L' is described b formula (L):

wherein L is a remainder of L';

A1 is a 1 -5 amino acid peptide covalently attached to the linking nitrogen in M1 or is absent; and A2 is a 1 -5 amino acid peptide covalently attached to the linking nitrogen in M2 or is absent.

3. The compound of claim 1 or 2, wherein the compound is described by formula (II):

(II)

wherein L is a remainder of L';

n is 1 , 2, 3, or 4;

each of R¹ , R¹², R'¹ , and R'¹² is, independently, a lipophilic moiety, a polar moiety, or H;

each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is, independently, optionally substituted C1 -C5 alkamino, a polar moiety, a positively charged moiety, or H;

each of R¹⁵ and R'¹⁵ is, independently, a lipophilic moiety or a polar moiety; each of R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R'², R'³, R'⁴, R'⁵, R'⁶, R'⁷, R'⁸, R'⁹, and R'¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; or two R groups on the same or adjacent atoms join to form an optionally substituted 5-8 membered ring;

each of a', b', c', a, b, and c is, independently, 0 or 1 ;

each of N¹ , N², N³, N⁴, N'¹ , N'², N'³, and N'⁴ is a nitrogen atom;

each of C¹ , C², C³, C'¹ , C'², and C'³ is a carbon atom,

or a pharmaceutically acceptable salt thereof.

4. The compound of claim 3, wherein the compound comprises at least one optionally substituted 5-8 membered ring formed by joining (i) R², R³, and C¹ ; (ii) R³, R⁴, N¹ , and C¹ ; (iii) R⁵, R⁶, and C²; (iv) R⁶, R⁷, N², and C²; (v) R⁸, R⁹, and C³; (vi) R⁹, R¹⁰, N³, and C³; (vii) R'², R'³, and C'¹ ; (viii) R'³, R'⁴, N'¹ , and C'¹ ; (ix) R'⁵, R'⁶, and C'²; (x) R'⁶, R'⁷, N'², and C'²; (xi) R'⁸, R'⁹, and C'³; or (xii) R'⁹, R'¹⁰, N'³, and C'³.

5. The compound of any one of claims 1 -4, wherein the compound is described by formula (III):

M2 A2 A1 M1

(III)

or a pharmaceutically acceptable salt thereof.

6. The compound of any one of claims 3-5, wherein

each of R¹ , R¹², R'¹ , and R'¹² is, independently, a lipophilic moiety;

each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is, independently, optionally substituted C1 -C5 alkamino, a polar moiety, or a positively charged moiety; and/or

each of R¹⁵ and R'¹⁵ is, independently, a polar moiety.

7. The compound of any one of claims 3-6, wherein each of R¹ and R¹² is a lipophilic moiety.

8. The compound of any one of claims 3-6, wherein each of R'¹ and R'¹² is a lipophilic moiety.

9. The compound of claim 7 or 8, wherein each lipophilic moiety is, independently, optionally substituted C1 -C20 alkyl, optionally substituted C5-C15 aryl, optionally substituted C6-C35 alkaryl, or optionally C5- C10 substituted heteroaryl.

10. The compound of any one of claims 7-9, wherein each lipophilic moiety is, independently, C1 -C8 alkyl, methyl substituted C2-C4 alkyl, (C1 -C10)alkylene(C6)aryl, phenyl substituted (C1 - C10)alkylene(C6)aryl, or alkyl substituted C4-C9 heteroaryl.

1 1 . The compound of any one of claims 7-10, wherein each lipophilic moiety is, independently, benzyl, isobutyl, sec-butyl, isopropyl, n-propyl, methyl, biphenylmethyl, n-octyl, or methyl substituted indolyl.

12. The compound of any one of claims 3-1 1 , wherein each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is independently optionally substituted C1 -C5 alkamino.

13. The compound of claim 12, wherein each of R^{1 1} , R¹³, R¹⁴, R'^{1 1} , R'¹³, and R'¹⁴ is CH2CH2NH2.

14. The compound of any one of claims 3-13, wherein each of R¹⁵ and R'¹⁵ is a polar moiety.

15. The compound of claim 14, wherein each polar moiety comprises a hydroxyl group, a carboxylic acid group, an ester group, or an amide group.

16. The compound of claim 14 or 15, wherein each polar moiety is hydroxyl substituted C1 -C4 alkyl.

17. The compound of any one of claims 14-16, wherein each polar moiety is CH(CH3)OH.

18. The compound of claim 3, wherein the compound is described by formula (IV):

(IV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl,

or a pharmaceutically acceptable salt thereof.

19. The compound of claim 18, wherein the compound is described by formula (IV-1 ):

(IV-1 )

or a pharmaceutically acceptable salt thereof.

20. The compound of claim 18, wherein the compound is described by formula (IV-2):

(IV-2) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl,

or a pharmaceutically acceptable salt thereof.

21 . The compound of claim 3, wherein the compound is described by formula (V):

(V)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl;

each of R², R⁶, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyi, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl,

or a pharmaceutically acceptable salt thereof.

22. The compound of claim 21 , wherein the compound is described by formula (V-1 ):

(V-1 )

or a pharmaceutically acceptable salt thereof.

23. The compound of claim 22, wherein the compound is described by formula (V-2):

(V-2)

or a pharmaceutically acceptable salt thereof.

24. The compound of claim 21 , wherein the compound is described by formula (V-3):

(V-3)

or a pharmaceutically acceptable salt thereof.

25. The compound of claim 24, wherein the compound is described by formula (V-4):

(V-4) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl;

or a pharmaceutically acceptable salt thereof.

26. The compound of claim 24 or 25, wherein the compound is described by formula (V-5):

(V-5)

or a pharmaceutically acceptable salt thereof.

27. The compound of claim 3, wherein the compound is described by formula (VI):

(VI)

each of R², R⁶, R⁸, R'², R'⁶, and R'⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyi, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl,

or a pharmaceutically acceptable salt thereof.

28. The compound of claim 27, wherein the compound is described by formula (VI-1 ):

(VI-1 )

or a pharmaceutically acceptable salt thereof.

29. The compound of claim 28, wherein the compound is described by formula (VI-2):

(VI-2)

or a pharmaceutically acceptable salt thereof.

30. The compound of claim 27, wherein the compound is described by formula (VI-3):

(VI-3) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl;

or a pharmaceutically acceptable salt thereof.

31 . The compound of claim 30, wherein the compound is described by formula (VI-4):

(VI-4)

or a pharmaceutically acceptable salt thereof.

32. The compound of claim 30 or 31 , wherein the compound is described by formula (VI-5):

(VI-5)

or a pharmaceutically acceptable salt thereof.

33. The compound of claim 3, wherein the compound is described by formula (VII):

(VII)

each of R², R⁶, R⁸, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl,

or a pharmaceutically acceptable salt thereof.

34. The compound of claim 3, wherein the compound is described by formula (VIII):

(VIM)

each of R², R⁶, R⁸, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl,

or a pharmaceutically acceptable salt thereof.

35. The compound of claim 3, wherein the compound is described by formula (IX):

(IX)

each of R², R⁶, and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl,

or a pharmaceutically acceptable salt thereof.

36. The compound of claim 35, wherein the compound is described by formula (IX-1 ) :

(IX-1 )

or a pharmaceutically acceptable salt thereof.

37. The compound of claim 36, wherein the compound is described by formula (IX-2):

(IX-2)

or a pharmaceutically acceptable salt thereof.

38. The compound of claim 3, wherein the compound is described by formula (X):

(X)

each of R² and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl,

or a pharmaceutically acceptable salt thereof.

39. The compound of claim 3, wherein the compound is described by formula (XI):

(XI)

each of R² and R⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl,

or a pharmaceutically acceptable salt thereof.

40. The compound of claim 39, wherein the compound is described by formula (XI-1 ) :

(XI-1 )

or a pharmaceutically acceptable salt thereof.

41 . The compound of claim 40, wherein the compound is described by formula (XI-2):

(XI-2)

or a pharmaceutically acceptable salt thereof.

The compound of claim 1 or 2, wherein the compound is described by formula (XII):

each of R² and R'² is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

R⁶, R⁷, N², and C² together form an optionally substituted 5-8 membered ring comprising optionally substituted C3-C7 heterocycloalkyl comprising an N heteroatom and additional 0-2

heteroatoms independently selected from N, O, and S, or optionally substituted C2-C7 heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S; R'⁶, R'⁷, N'², and C'² together form an optionally substituted 5-8 membered ring comprising optionally substituted C3-C7 heterocycloalkyl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S, or optionally substituted C2-C7 heteroaryl comprising an N heteroatom and additional 0-2 heteroatoms independently selected from N, O, and S;

or a pharmaceutically acceptable salt thereof.

43. The compound of claim 42, wherein the compound is described by formula (XI 1-1 ):

(XII-1 )

or a pharmaceutically acceptable salt thereof.

44. The compound of claim 1 or 2, wherein the compound described by formula (XIII):

E'¹ E¹

L !'i¹ L Ί¹

I I

M2— A2— L A1 M1

(XIII)

wherein A1 is a 1 -5 amino acid peptide covalently attached to the linking nitrogen in M1 ;

A2 is a 1 -5 amino acid peptide covalently attached to the linking nitrogen in M2;

E¹ is a first monosaccharide or oligosaccharide moiety;

E'¹ is a second monosaccharide or oligosaccharide moiety;

L, L¹ , and L'¹ are remainders of L';

or a pharmaceutically acceptable salt thereof.

45. The compound of claim 44, wherein the compound is described by formula (XIII-1 ):

(XIII-1 )

each of R⁶ and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each of R² and R'² is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2- C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene;

or a pharmaceutically acceptable salt thereof.

46. The compound of claim 45, wherein the compound is described by formula (XI 11-2) :

(XIII-2)

or a pharmaceutically acceptable salt thereof.

47. The compound of claim 46, wherein the compound is described by formula (XIII-3):

(XIII-3)

or a pharmaceutically acceptable salt thereof.

48. The compound of claim 1 or 2, wherein the compound described by formula (XIV):

E

L

M2— A2— L A1 M1

(XIV)

A2 is a 1 -5 amino acid peptide covalently attached to the linking nitrogen in M2, or is absent; L and L¹ are remainders of L';

or a pharmaceutically acceptable salt thereof.

49. The compound of any one of claims 21 , 27, 33-35, 38, 39, and 42, wherein R² is optionally substituted C1 -C5 alkamino.

50. The compound of any one of claims 21 , 27, 33-35, 38, and 42, wherein R'² is optionally substituted C1 -C5 alkamino.

51 . The compound of claim 49 or 50, wherein the optionally substituted C1 -C5 alkamino is CH2NH2 or

52. The compound of any one of claims 21 , 27, 33-35, 38, 39, and 42, wherein R² is a polar moiety.

53. The compound of any one of claims 21 , 27, 33-35, 38, and 42, wherein R'² is a polar moiety.

54. The compound of claim 52 or 53, wherein the polar moiety comprises a hydroxyl group, a carboxylic acid group, an ester group, or an amide group.

55. The compound of any one of claims 52-54, wherein the polar moiety is hydroxyl substituted C1 -C4 alkyl.

56. The compound of claim 55, wherein the polar moiety is CH(CH3)OH or CH2OH.

57. The compound of any one of claims 21 , 27, 33-35, 39, and 45, wherein R⁶ is a polar moiety.

58. The compound of any one of claims 21 , 27, 33, and 45, wherein R'⁶ is a polar moiety.

59. The compound of claim 57 or 58, wherein the polar moiety comprises a hydroxyl group, a carboxylic acid group, an ester group, or an amide group.

60. The compound of any one of claims 57-59, wherein the polar moiety is hydroxyl substituted C1 -C4 alkyl.

61 . The compound of claim 60, wherein the polar moiety is CH(CH3)OH or CH2OH.

62. The compound of any one of claims 27, 33, and 34, wherein R⁸ is optionally substituted C1 -C5 alkamino.

63. The compound of claim 27, wherein R'⁸ is optionally substituted C1 -C5 alkamino.

64. The compound of claim 62 or 63, wherein the optionally substituted C1 -C5 alkamino is CH2NH2 or

65. The compound of any one of claims 27, 33, and 34, wherein R⁸ is optionally substituted C5-C1 5 aryl.

66. The compound of claim 27, wherein R'⁸ is optionally substituted C5-C15 aryl.

67. The compound of claim 65 or 66, wherein the optionally substituted C5-C1 5 aryl is naphthyl.

68. The compound of claim 42, wherein R⁶, R⁷, N², and C² together form a 5- or 6-membered ring comprising C4-C5 heterocycloalkyl comprising an N heteroatom and additional 0 or 1 heteroatom independently selected from N, O, and S; and wherein R'⁶, R'⁷, N'², and C'² together form a 5- or 6- membered ring comprising C4-C5 heterocycloalkyl comprising an N heteroatom and additional 0 or 1 heteroatom independently selected from N, O, and S.

69. The compound of claim 45, wherein each of R² and R'² is C1 -C4 heteroalkylene.

70. The compound of claim 69, wherein each of R² and R'² is -(CH2)2NH- or -CH2NH-.

The compound of claim 3, wherein the compound is described by formula (XV):

(XV)

or a pharmaceutically acceptable salt thereof.

72. The compound of any one of claims 1 -71 , wherein L', L, L¹ , or L'¹ comprises one or more optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, optionally substituted C2-C15 heteroarylene, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino, wherein R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl.

73. The compound of claims 72, wherein the backbone of L', L, L¹ , or L'¹ consists of one or more optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, optionally substituted C2-C15 heteroarylene, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino; and wherein R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C1 5 heteroaryl.

74. The compound of 72 or 73, wherein L', L, L¹ , or L'¹ is oxo substituted.

75. The compound of any one of claims 1 -74, wherein the backbone of L', L, L¹ , or L'¹ comprises no more than 250 atoms.

76. The compound of any one of claims 1 -75, wherein L', L, L¹ , or L'¹ is capable of forming an amide, a carbamate, a sulfonyl, or a urea linkage.

77. The compound of any one of claims 2-71 , wherein L, L¹ , or L'¹ is a bond.

78. The compound of any one of claims 2-43 and 71 , wherein L is described by formula (L-l):

L^c

L^B-Q-L^A (L-l)

G^B2;

G^C2;

G^A1 is a bond attached to Q in formula (L-l);

G^A2 is a bond attached to A1 or M1 if A1 is absent;

G^B1 is a bond attached to Q in formula (L-l);

G^B2 is a bond attached to A2 or M2 if A2 is absent;

G^C1 is a bond attached to Q in formula (L-l);

G^C2 is a bond attached to E;

each of Z^A1 , Z^A2, Z^A3, Z^M, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4 and Z^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene;

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, and o3 is, independently, 0 or 1 ;

Q is a nitrogen atom, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20

heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene.

441

wherein R^* is a bond or comprises one or more of optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, optionally substituted C2-C15

heteroarylene, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, and imino, and

80. The compound of any one of claims 2-43 and 71 , wherein L is described by formula (L-lla) or formula (L-llb):

(L-lla) (L-llb)

L^B is described by formula G^B1-(Z^B1 )_g2-(Y^B1 )h2-(Z^B2)i2-(Y^B2)j2-(Z^B3)_k2-(Y^B3)i2-(Z^B4)m2-(Y^B4)n2-(Z^B5)o2-

G^B2;

L^c is described by formula G^c1 -(Z^c1 )_g3-(Y^c1 )h3-(Z^C2)i₃-(Y^C2)j3-(Z^C3)k3-(Y^C3)i3-(Z^C4)m3-

G^D2;

G^A1 is a bond attached to N^A in formula (L-lla) or N^A in formula (L-llb);

G^A2 is a bond attached to A1 or M1 if A1 is absent;

G^B1 is a bond attached to N^A in formula (L-lla) or N^A in formula (L-llb);

G^B2 is a bond attached to A2 or M2 if A2 is absent;

G^C1 is a bond attached to N^B in formula (L-lla) or C in formula (L-llb);

G^D1 is a bond attached to N^B in formula (L-lla) or C in formula (L-llb);

G^D2 is a bond attached to a second monosaccharide or oligosaccharide moiety, E'¹ ;

each of Z^A1 Z^A2 Z^A3 Z^A4 Z^A5 Z^B1 Z^B2 Z^B3 Z^B4 Z^B5 Z^C1 Z^C2 Z^C3 Z^C4 Z^C5 Z^D1 Z^D2 Z^D3 Z^D4 and Z^D5 is independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene;

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; and each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, o3, g4, h4, i4, j4, k4, I4, m4, n4, and o4 is, independently, 0 or 1 . The compound of claim 80, wherein L is

82. The compound of any one of claims 44-47, wherein L is described by formula (L-lll):

H^A1 -(W^A1 )_gi -(X^A1 )hi -(W^A2)ii -(X^A2)ji -(W^A3)ki -(X^A3)ii -(W^A4)_mi -(X^A4)m -(W^A5)₀1 -H^A2

(L-lll)

wherein H^A1 is a bond attached to A2;

H^A2 is a bond attached to A1 ;

each of W^A1 , W^A2, W^A3, W^M , and W^A5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene;

each of X^A1 , X^A2, X^A3, X^A4 is, independently, O, S, NR\ P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino;

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; and each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , and o1 is, independently, 0 or 1 .

83. The compound of claim 82, where L is

84. The compound of any one of claims 44-47, 82, and 83, wherein L¹ is described by formula (L-IV):

H^B1-(W^B1 )g2-(X^B1 )h2-(W^B2)i2-(X^B2)j2-(W^B3)k2-(X^B3)l2-(W^B4)m2-(X^B4)n2-(W^B5)o2-H

(L-IV)

wherein H^B1 is a bond attached to A1 ;

H^B2 is a bond attached to a first monosaccharide or oligosaccharide moiety, E¹ ;

each of W^B1 , W^B2, W^B3, W^B4, and W^B5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene;

each of X^B1 , X^B2, X^B3, X^B4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino;

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; and each of g2, h2, i2, j2, k2, I2, m2, n2, and o2 is, independently, 0 or 1 .

86. The compound of any one of claims 44-47 and 82-85, wherein L'¹ is described by formula (L-V):

H^C1 -(W^C1 )g₃-(X^C1 )h3-(W^C2)_i3-(X^C2)j3-(W^C3)k3-(X^C3)l3-(W^C )m3-(X^C4)n3-(W^C5)o3H^C2

(L-V)

wherein H^C1 is a bond attached to A2;

H^C2 is a bond attached to a second monosaccharide or oligosaccharide moiety, E'¹ ;

each of W^C1 , W^C2, W^C3, W^C4, and W^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-

C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C15 arylene, or optionally substituted C2-C15 heteroarylene;

each of X^C1 , X^C2, X^C3, X^C4 is, independently, O, S, NR', P, carbonyl, thiocarbonyl, sulfonyl, phosphate, phosphoryl, or imino;

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ; and each of g3, h3, i3, j3, k3, I3, m3, n3, and o3 is, independently, 0 or 1 .

90. The compound of any one of claims 1 -87 or 342-368, wherein E, E¹ , or E'¹ is any one of the moieties in Tables 2A and 2B.

91 . The compound of any one of claims 1 -87 or 342-368, wherein E, E¹ , or E'¹ directly or indirectly activates an immune cell.

92. The compound of any one of claims 1 -87 or 342-368, wherein E, E¹ , or E'¹ is a ligand to an innate immune receptor.

93. The compound of claim 92, wherein the innate immune receptor is AICL, BDCA2, CLEC2,

Complement receptor 3, Complement receptor 4, DCIR, dectin-1 , dectin-2, DC-SIGN, a C-Type lectin receptor, MMR, langerin, TLR2, Mincle, MBL, or KCR.

94. The compound of claims 1 -87 or 342-368, wherein E, E¹ , or E'¹ binds to an antibody.

95. The compound of claim 94, wherein the antibody is a natural antibody.

96. The compound of claim 95, wherein the natural antibody is an antibody of the immunoglobulin M (IgM) isotype.

97. The compound of any one of claims 94-96, wherein the antibody binds to a moiety in Tables 2A and 2B.

98. The compound of any one of claims 94-97, wherein the antibody is anti-aGal antibody or anti-aRha antibody.

99. The compound of any one of claims 1 -87 or 342-368, wherein the monosaccharide moiety has one optionally substituted C6-C9 monosaccharide residue.

100. The compound of any one of claims 1 -87 or 342-368, wherein the oligosaccharide moiety has 2-18 optionally substituted C6-C9 monosaccharide residues.

101 . The compound of claim 100, wherein the oligosaccharide moiety has 2-12 optionally substituted C6- C9 monosaccharide residues.

102. The compound of any one of claims 99-101 , wherein each of the optionally substituted C6-C9 monosaccharide residues is, independently, glucose (Glc), galactose (Gal), mannose (Man), allose (All), altrose (Alt), gulose (Gul), idose (Ido), talose (Tal), fucose (Fuc), rhamnose (Rha or L-Rha), thia- rhamnose (thia-Rha or thia-L-Rha), quinovose (Qui), 2-deoxyglucose (2-dGlc), glucosamine (GlcN), galactosamine (GaIN), mannosamine (ManN), fucosamine (FucN), quinovosamine (QuiN), N-Acetyl- glucosamine (GlcNAc), N-Acetyl-galactosamine (GalNAc), N-Acetyl-mannosamine (ManNAc), N-acetyl- fucosamine (FucNAc), N-acetyl-quinovosamine (QuiNAc), glucuronic acid (GlcA), galacturonic acid (GalA), mannuronic acid (ManA), iduronic acid (IdoA), sialic acid (Sia), neuraminic acid (Neu), N-Acetyl- neuraminic acid (Neu5Ac), N-Glycolyl-neuraminic acid (Neu5Gc), glucitol (Glc-ol), galactitol (Gal-ol), mannitol (Man-ol), fructose (Fru), sorbose (Sor), tagatose (Tag), thevetose (The), acofriose (Aco), digitoxose (Dig), cymarose (Cym), abequose (Abe), colitose (Col), tyvelose (Tyv), ascarylose (Asc), paratose (Par), or N-acetyl-muramic acid (MurNAc).

103. The compound of claim 102, wherein each of the optionally substituted C6-C9 monosaccharide residues is, independently, an optionally substituted C6 monosaccharide residue.

104. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is Glc, Gal, Man, All, Alt, Gul, Ido, or Tal.

105. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is Fuc, Rha or L-Rha, thia-Rha or thia-L-Rha, Qui, or 2-dGlc.

106. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is GlcN, GaIN, ManN, FucN, or QuiN.

107. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is N- GlcNAc, GalNAc, ManNAc, FucNAc, QuiNAc, GlcA, GalA, ManA, or IdoA.

108. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is Glc- ol, Gal-ol, or Man-ol.

109. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is Fru, Sor, Tag.

1 10. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is The, Aco, Dig, Cym, Abe, Col, Tyv, Asc, Par, or MurNAc.

1 1 1 . The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is Rha, Gal, Glc, GlcA, GlcNAc, GalNAc, GlcN(Gc) (N-Glycolyl-Glucosamine), Neu5Ac, Neu5Gc, Fuc,

Man, -hbPOsMan (mannose phosphate), 6-H2PO3GIC (glucose phosphate), Mur (muramoyl), Mur-L-Ala-D- i-Gln-Lys, (Mur)-3-0-GlcNAc, sulfate-galactose (Su-Gal), disulfate-galactose (Su2-Gal), sulfate-glucose (Su-Glc), sulfate-GlcNAc (Su-GlcNAc), or sulfate-GalNAc (Su-GalNAc).

1 12. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is optionally substituted Rha.

1 13. The compound of claim 1 12, wherein the optionally substituted C6 monosaccharide residue is Rha.

1 14. The compound of claim 1 12 or 1 13, wherein the optionally substituted C6 monosaccharide residue is L-Rha.

1 15. The compound of claim 103, wherein the optionally substituted C6 monosaccharide residue is optionally substituted Gal or optionally substituted Glc.

1 16. The compound of claim 103, wherein the optionally substituted Gal is optionally substituted a1 -3Gal.

1 17. The compound of claim 103, wherein the optionally substituted Glc is an optionally substituted β- glucan having 1 -6 Glc moieties.

1 18. The compound of claim 1 17, wherein the optionally substituted β-glucan is

pustulan.

1 19. The compound of claim 1 18, wherein the optionally substituted β-glucan is laminarin.

120. The compound of claim 102, wherein each of the optionally substituted C6-C9 monosaccharide residues is, independently, an optionally substituted C9 monosaccharide residue, wherein the optionally substituted C9 monosaccharide residue is Sia, Neu, Neu5Ac, or Neu5Gc.

121 . The compound of any one of claims 102-120, wherein at least one optionally substituted C6-C9 monosaccharide residue is substituted with 1 -3 substituents independently selected from sulfate, phosphate, methyl, acetyl, natural amino acids, and non-natural amino acids.

122. The compound of any one of claims 102-120, wherein at least one optionally substituted C6-C9 monosaccharide residue is substituted with 1 -3 substituents independently selected from sulfate, phosphate, methyl, and acetyl.

123. The compound of any one of claims 102-120, wherein the at least one optionally substituted C6-C9 monosaccharide moiety is substituted with 1 -3 substituents independently selected from natural and non- natural amino acids.

124. The compound of claim 123, wherein the natural amino acid is alanine, lysine, serine, glutamine or asparagine.

125. A compound of formula (XVI):

(XVI)

wherein:

each A is an independently selected amino acid;

L is a linker that, when m is 2, 3, 4, or 5, is bound to any of A;

each E is independently selected from a monosaccharide or an oligosaccharide;

m is 0, 1 , 2, 3, 4, or 5;

n is 1 , 2, 3, 4, or 5; and

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; or a pharmaceutically acceptable salt thereof.

126. The compound of claim 125, wherein Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of a natural amino acid, or a pharmaceutically acceptable salt thereof.

127. The compound of claim 125, wherein at least one of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

128. The compound of claim 125, wherein at least two of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

129. The compound of claim 125, wherein at least three of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

130. The compound of claim 125, wherein at least four of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

131 . The compound of claim 125, wherein at least five of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

132. The compound of claim 125, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

133. The compound of claim 125, wherein each Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of serine, threonine, cysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan, or a pharmaceutically acceptable salt thereof.

134. The compound of claim 125, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyl, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

135. The compound of claim 134, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

136. The compound of claim 135, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

A compound of formula (XVII):

(XVII)

wherein:

each A¹ and A² is an independently selected amino acid;

L is a linker that, when m is 1 , 2, 3, 4, or 5, is bound to a nitrogen atom in any A¹ and a nitrogen atom in any A²;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

n is 1 , 2, 3, 4, or 5; and

138. The compound of claim 137, wherein Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of a natural amino acid, or a pharmaceutically acceptable salt thereof.

139. The compound of claim 137, wherein at least one of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

140. The compound of claim 137, wherein at least two of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

141 . The compound of claim 137, wherein at least three of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

142. The compound of claim 137, wherein at least four of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

143. The compound of claim 137, wherein at least five of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

144. The compound of claim 137, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of a non-natural amino acid, or a pharmaceutically acceptable salt thereof.

145. The compound of claim 137, wherein each Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from the side chain of serine, threonine, cysteine, glycine, proline, alanine, valine, isoleucine, leucine, methionine, phenylalanine, tyrosine, and tryptophan, or a pharmaceutically acceptable salt thereof.

146. The compound of claim 137, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

147. The compound of claim 137, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

148. The compound of claim 137, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

149. The compound of any one of claims 137-148, wherein:

each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5- yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3-hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2- aminooctanoic acid, serine, 2-aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha-neopentylglycine; and

each m is independently selected from 1 , 2, 3, 4, or 5;

or a pharmaceutically acceptable salt thereof.

150. The compound of any one of claims 137-149, wherein each m is independently 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

151 . The compound of any one of claims 137-150, wherein n is 1 , 2, 3, or 4, or a pharmaceutically acceptable salt thereof.

152. The compound of any one of claims 137-151 , wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

153. The compound of claim 137, wherein:

each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H- tetrazol-5-yl)alanine, 3-aminoalanine, piperazine-2-carboxylic acid, 2,4-diaminobutyric acid, 3- hydroxyproline, threonine, 2-amino-4-phenylbutyric acid, 3-(2-naphthyl)alanine, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, serine, 2-aminohexanoic acid, 4-amino-4-piperidinyl carboxylic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S- propylhomocysteine, cyclopropylalanine, 3-fluoroalanine, 2-amino-5-methylhexanoic acid, 2-amino-5- methylhex-4-enoic acid, alpha-t-butylglycine, and alpha-neopentylglycine;

each m is independently 1 , 2, 3 or 4;

E is a monosaccharide;

n is 1 , 2, 3, or 4; and

the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

A compound of formula (XVIII):

(XVIII)

wherein:

each A¹ and A² is an independently selected amino acid; X is absent or is -CH₂CH₂C(0)NH-;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

n is 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 10; and

e is an integer from 0 to 10;

or a pharmaceutically acceptable salt thereof.

155. The compound of claim 154, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

156. The compound of claim 155, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

157. The compound of claim 154, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

158. The compound of claim 157, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

159. The compound of any one of claims 154-158, wherein:

m is 2 or 3;

d is an integer from 1 to 10;

e is an integer from 1 to 10; and E is a monosaccharide;

or a pharmaceutically acceptable salt thereof.

160. The compound of claim 154, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; m is 2 or 3;

d is an integer from 1 to 10;

e is an integer from 1 to 10;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

161 . The compound of claim 160, wherein m is 2, or a pharmaceutically acceptable salt thereof. 162. The compound of claim 160, wherein m is 3, or a pharmaceutically acceptable salt thereof. 163. The compound of claim 160, wherein:

d is 10;

e is 10; and

X is absent;

or a pharmaceutically acceptable salt thereof.

164. The compound of claim 160, wherein:

m is 2;

d is 1 ;

e is 1 ; and

X is -CH₂CH₂C(0)NH-;

or a pharmaceutically acceptable salt thereof.

165. The compound of any one of claims 154-164, wherein

E is

or a pharmaceutically acceptable salt thereof.

166. The compound of claim 165, wherein E is OH , or a pharmaceutically acceptable thereof.

A compound of formula (XIX):

(XIX)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

n is 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 10;

e is an integer from 0 to 10;

f is an integer from 0 to 10; and

g is an integer from 0 to 10;

or a pharmaceutically acceptable salt thereof.

168. The compound of claim 167, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

169. The compound of claim 168, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

170. The compound of claim 167, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceutca y accepta e sat t ereo .

171. The compound of claim 170, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

172. The compound of any one of claims 167-171 , wherein:

m is 2;

d is 1 ;

e is 1 ;

f is 3;

g is 1 ; and

E is a monosaccharide;

or a pharmaceutically acceptable salt thereof.

173. The compound of claim 167, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine;

m is 2;

d is 1 ;

e is 1 ;

f is 3;

g is 1 ;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

1 74. The compound of any one of claims 1 67-1 73, wherein

or a pharmaceutically acceptable salt thereof.

1 75. The compound of claim 1 74, wherein E is

, or a pharmaceutically acceptable salt thereof.

A compound of formula (XX) :

(XX)

wherein :

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 1 0;

e is an integer from 0 to 1 0;

f is an integer from 0 to 1 0; and

g is an integer from 0 to 1 0;

or a pharmaceutically acceptable salt thereof.

1 77. The compound of claim 1 76, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

178. The compound of claim 177, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

179. The compound of claim 178, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

180. The compound of claim 179, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

181 . The compound of any one of claims 176-180, wherein:

m is 2;

d is 1 ;

e is 1 ;

f is 1 ;

g is 1 ; and

E is a monosaccharide;

or a pharmaceutically acceptable salt thereof.

182. The compound of claim 176, wherein:

m is 2;

d is 1 ;

e is 1 ;

f is 1 ;

g is 1 ;

E is a monosaccharide; and

n is 1 ; or a pharmaceutically acceptable salt thereof.

183. The compound of any one of claims 176-182, wherein

or a pharmaceutically acceptable salt thereof.

184. e compoun o c a m 183, w ere n s , or a p armaceut ca y accepta e thereof.

185. A compound of formula (XXI):

(XXI)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

e is an integer from 0 to 10;

f is an integer from 0 to 10;

g is an integer from 0 to 25;

or a pharmaceutically acceptable salt thereof.

186. The compound of claim 185, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

187. The compound of claim 186, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

188. The compound of claim 187, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

189. The compound of claim 188, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

190. The compound of any one of claims 185-189, wherein:

m is 2;

d is 1 ;

e is 1 ;

f is 1 ;

g is 8 to 25; and

E is a monosaccharide;

or a pharmaceutically acceptable salt thereof.

191 . The compound of claim 185, wherein:

m is 2;

d is 1 ;

e is 1 ;

f is 1 ;

g is 8 to 25; E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

192. The compound of any one of claims 185-191 , wherein

or a pharmaceutically acceptable salt thereof.

193. The compound of claim 192, wherein E is

, or a pharmaceutically acceptable salt thereof.

A compound of formula (XXII):

(XXII)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; and

d is an integer from 0 to 15;

or a pharmaceutically acceptable salt thereof.

195. The compound of claim 194, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

196. The compound of claim 195, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

197. The compound of claim 196, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

198. The compound of claim 197, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

199. The compound of any one of claims 194-198, wherein:

m is 2;

d is 10;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

200. The compound of claim 194, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid and piperazine-2- carboxylic acid;

m is 2;

d is 10;

E is a monosaccharide; and n is 1 ;

or a pharmaceutically acceptable salt thereof.

201 . The compound of any one of claims 194-200, wherein

or a pharmaceutically acceptable salt thereof.

202. The compound of claim 201 , wherein E is

, or a pharmaceutically acceptable salt thereof.

A compound of formula (XXIII):

(XXIII)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

Y is

, -C(0)CH₂CH₂-, -CH2-, or is absent;

X is -C(0)CH₂CH2- or is absent;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 15; e is an integer from 1 to 10;

f is an integer from 1 to 5; and

g is an integer from 1 to 5;

or a pharmaceutically acceptable salt thereof.

204. The compound of claim 203, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

205. The compound of claim 204, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

206. The compound of claim 205, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

207. The compound of claim 206, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

208. The compound of any one of claims 203-207, wherein:

m is 2, 3 or 4;

d is 1 ;

E is a monosaccharide;

f is 1 or 2; g is 1 or 2; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

209. The compound of claim 203, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 3-aminoalanine, 2- piperazinecarboxylic acid, 2-aminohexanoic acid, 2-aminooctanoic acid, methionine, and threonine; m is 2, 3 or 4;

d is 1 ;

E is a monosaccharide;

f is 1 or 2;

g is 1 or 2; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

210. The compound of any one of claims 203-207, wherein:

m is 3 or 4;

d is 1 ;

Y is absent;

E is a monosaccharide;

f is 1 ;

g is 1 ; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

21 1 . The compound of claim 210, wherein

each A¹ and A² is independently selected from 2,4-diaminobutyric acid and threonine; and m is 3;

or a pharmaceutically acceptable salt thereof.

212. The compound of any one of claims 203-207, wherein:

m is 2, 3 or 4;

d is 1 ;

Y is -C(0)CH₂CH₂-;

E is a monosaccharide;

f is 1 or 2;

g is 1 or 2; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

213. The compound of claim 212, wherein each A¹ and A² is independently selected from glycine, arginine, asparagine, glutamine, 3-(2H-tetrazol-5-yl)alanine, 2,4-diaminobutyric acid, 3-aminoalanine, 2- aminohexanoic acid, 2-piperazinecarboxylic acid, 2-aminooctanoic acid, methionine, methionine sulfoxide, methionine sulfone, S-methylcysteine, S-ethylcysteine, S-propylhomocysteine,

cyclopropylalanine, 3-fluoroalanine, 2-amino-5-methylhexanoic acid, 2-amino-5-methylhex-4-enoic acid, alpha-t-butylglycine, and alpha-neopentylglycine, or a pharmaceutically acceptable salt thereof.

214. The compound of claim 213, wherein m is 4, d is 1 , f is 1 , and g is 1 , or a pharmaceutically acceptable salt thereof.

215. The compound of claim 213, wherein m is 3, f is 2, and g is 1 , or a pharmaceutically acceptable salt thereof.

216. The compound of any one of claims 203-207, wherein:

m is 2, 3 or 4;

d is 1 ; E is a monosaccharide;

f is 1 or 2;

g is 1 or 2; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

217. The compound of claim 216, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid and threonine;

f is 1 ; and

g is 1 ;

or a pharmaceutically acceptable salt thereof.

218. The compound of claim 216, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminohexanoic acid, 2- aminooctanoic acid and threonine, or a pharmaceutically acceptable salt thereof;

m is 4;

f is 1 ; and

g is 1 ;

or a pharmaceutically acceptable salt thereof.

219. The compound of claim 217, wherein:

m is 3;

f is 1 ; and

g is 1 ;

or a pharmaceutically acceptable salt thereof.

220. The compound of any one of claims 203-219, wherein

E is

or a pharmaceutically acceptable salt thereof.

221 . The compound of claim 220, wherein E is OH , or a pharmaceutically acceptable thereof.

222. A compound of formula (XXIV):

(XXIV)

wherein:

each A¹ and A² is an independently selected amino acid;

X is -C(0)CH₂CH₂CH2-Y- or -C(0)CH₂CH₂C(0)NH-;

Y is heteroaryl ;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

d is an integer from 0 to 15;

or a pharmaceutically acceptable salt thereof.

223. The compound of claim 222, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

224. The compound of claim 223, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl. hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

225. The compound of claim 224, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

226. The compound of claim 225, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

227. The compound of any one of claims 222-226, wherein:

m is 2 or 3;

d is an integer from 1 to 3;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

228. The compound of claim 227, wherein each A¹ and A² is independently selected from 2,4- diaminobutyric acid, 3-(2-naphthyl)alanine, and threonine, or a pharmaceutically acceptable salt thereof.

229. The compound of claim 228, wherein:

X is -C(0)CH₂CH₂CH2-Y-;

m is 3;

d is 3; and

Y is 1 ,4-triazololyl;

or a pharmaceutically acceptable salt thereof.

230. The compound of claim 228, wherein:

X is -C(0)CH₂CH₂C(0)-;

m is 2; and

d is 1 ;

or a pharmaceutically acceptable salt thereof.

31 . The compound of any one of claims 222-230, wherein

or a pharmaceutically acceptable salt thereof.

232. The compound of claim 228, wherein E is

, or a pharmaceutically acceptable salt thereof.

A compound of formula (XXV):

(XXV)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 15; and

e is an integer from 0 to 15;

or a pharmaceutically acceptable salt thereof.

234. The compound of claim 233, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

235. The compound of claim 234, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

236. The compound of claim 235, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

237. The compound of claim 236, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

238. The compound of any one of claims 233-237, wherein:

m is 2 or 3;

d is an integer from 1 to 3;

e is an integer from 1 to 3;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

239. The compound of claim 233, wherein:

m is 2 or 3;

d is an integer from 1 to 3;

e is an integer from 1 to 3;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

240. The compound of any one of claims 233-239, wherein

E is

;

or a pharmaceutically acceptable salt thereof.

241 . The compound of claim 240, wherein E is

, or a pharmaceutically acceptable salt thereof.

A compound of formula (XXVI):

(XXVI)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

R is C1 -C20 alkyl;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 20;

e is an integer from 0 to 20; and

f is an integer from 0 to 20;

or a pharmaceutically acceptable salt thereof.

243. The compound of claim 242, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

244. The compound of claim 243, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

245. The compound of claim 244, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

246. The compound of claim 245, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

247. The compound of any one of claims 242-246, wherein:

m is 2 or 3;

d is an integer from 1 to 3;

e is an integer from 1 to 3;

f is an integer from 1 to 3;

E is a monosaccharide;

R is C1 -C10 alkyl; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

248. The compound of claim 242, wherein:

m is 2;

d is 1 ;

e is 1 ;

f is 1 ; E is a monosaccharide;

R is C1 -C10 alkyl; and

n is 1 ;

or a pharmaceutically acceptable salt thereof. 249. The compound of any one of claims 242-248, wherein

E is

or a pharmaceutically acceptable salt thereof.

250. The compound of claim 249, wherein E is

, or a pharmaceutically acceptable salt thereof.

251 . A compound of formula (XXVII):

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

R is C1 -C20 alkyl;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 20;

e is an integer from 1 to 20; and

f is an integer from 1 to 20;

or a pharmaceutically acceptable salt thereof.

252. The compound of claim 251 , wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

253. The compound of claim 252, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl. hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

254. The compound of claim 253, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

255. The compound of claim 251 , wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

256. The compound of any one of claims 251 -255, wherein:

m is 2, 3, or 4;

d is an integer from 1 to 3;

e is an integer from 1 to 3;

f is an integer from 1 to 3;

E is a monosaccharide;

R is C1 -C10 alkyl; and

n is 1 ; or a pharmaceutically acceptable salt thereof.

257. The compound of claim 251 , wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine;

m is 2, 3, or 4;

d is 1 ;

e is 1 ;

f is 1 ;

E is a monosaccharide;

R is C1 -C10 alkyl; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

258. The compound of claim 257, wherein m is 2, or a pharmaceutically acceptable salt thereof.

259. The compound of claim 257, wherein m is 3, or a pharmaceutically acceptable salt thereof.

260. The compound of claim 257, wherein m is 4, or a pharmaceutically acceptable salt thereof.

261 . The compound of any one of claims 251 -260, wherein

E is

or a pharmaceutically acceptable salt thereof.

262. The compound of claim 261 , wherein E is

, or a pharmaceutically acceptable salt thereof.

263. A compound of formula (XXVIII):

(XXVIII)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

Y is

, -C(0)CH₂CH₂-, -CH2-, or is absent;

e is an integer from 1 to 10;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

d is an integer from 0 to 15;

or a pharmaceutically acceptable salt thereof.

264. The compound of claim 263, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

265. The compound of claim 264, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

266. The compound of claim 265, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

267. The compound of claim 266, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

Q¹ Q² Q³ Q⁴ Q⁵ Q⁶

* 3 X Jb X Jb

or a pharmaceutically acceptable salt thereof.

268. The compound of any one of claims 263-267, wherein Y is -C(0)CH2CH2-, or a pharmaceutically acceptable salt thereof.

269. The compound of any one of claims 263-268, wherein:

m is 2, 3, or 4;

d is an integer from 1 to 3;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

270. The compound of claim 269, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; and

d is 1 ;

or a pharmaceutically acceptable salt thereof.

271 . The compound of claim 270, wherein m is 2, or a pharmaceutically acceptable salt thereof.

272. The compound of claim 270, wherein m is 3, or a pharmaceutically acceptable salt thereof.

273. The compound of claim 270, wherein m is 4, or a pharmaceutically acceptable salt thereof.

274. The compound of any one of claims 263-273, wherein

or a pharmaceutically acceptable salt thereof.

275. The compound of claim 273, wherein E is OH , or a pharmaceutically acceptable thereof.

A compound of formula (XXIX):

(XXIX)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

d is an integer from 0 to 15;

or a pharmaceutically acceptable salt thereof.

277. The compound of claim 276, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

278. The compound of claim 277, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl. hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

279. The compound of claim 278, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

280. The compound of claim 279, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

281 . The compound of any one of claims 276-280, wherein:

m is 2, 3, or 4;

d is an integer from 1 to 3;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

282. The compound of claim 281 , wherein:

d is 1 ;

or a pharmaceutically acceptable salt thereof.

283. The compound of claim 282, wherein m is 2, or a pharmaceutically acceptable salt thereof.

284. The compound of claim 282, wherein m is 3, or a pharmaceutically acceptable salt thereof.

285. The compound of claim 282, wherein m is 4, or a pharmaceutically acceptable salt thereof.

286. The compound of any one of claims 276-285, wherein

E is

;

or a pharmaceutically acceptable salt thereof.

287. The compound of claim 286, wherein E is

, or a pharmaceutically acceptable salt thereof.

A compound of formula (XXX):

(XXX)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

Y is

-C(0)CH₂CH₂-, -CH2-, or is absent;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 0 to 15;

g is an integer from 0 to 15; and

e and e' are independently an integer from 1 to 3;

or a pharmaceutically acceptable salt thereof.

289. The compound of claim 288, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

290. The compound of claim 289, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

291 . The compound of claim 290, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

292. The compound of claim 291 , wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

293. The compound of any one of claims 288-292, wherein Y is -C(0)CH2CH2-, or a pharmaceutically acceptable salt thereof.

294. The compound of any one of claims 288-293, wherein:

m is 2, 3, or 4;

d is an integer from 1 to 3;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

295. The compound of claim 294, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminooctanoic acid, and threonine; and d is 1 ; and

e is 1 or 2;

or a pharmaceutically acceptable salt thereof.

296. The compound of claim 295, wherein m is 2, or a pharmaceutically acceptable salt thereof.

297. The compound of claim 295 wherein m is 3, or a pharmaceutically acceptable salt thereof.

298. The compound of claim 295, wherein m is 4, or a pharmaceutically acceptable salt thereof.

299. The compound of any one of claims 288-298, wherein

or a pharmaceutically acceptable salt thereof.

300. The compound of claim 299, wherein E is

, or a pharmaceutically acceptable salt thereof.

301 . A compound of formula (XXXI):

(XXXI)

wherein: each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each Y is independently selected from

, -C(0)CH2CH2-, and -CH2-, or is absent; each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 15;

each e is independently from 1 to 15;

each f is independently from 1 to 15; and

g is from 1 to 15;

or a pharmaceutically acceptable salt thereof.

302. The compound of claim 301 , wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

303. The compound of claim 302, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

304. The compound of claim 303, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

305. The compound of any one of claims 301 -304, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

306. The compound of any one of claims 301 -305, wherein Y is -C(0)CH2CH2-, or a pharmaceutically acceptable salt thereof.

307. The compound of any one of claims 301 -306, wherein:

m is 2, 3, or 4;

d is an integer from 1 to 5;

e is an integer from 1 to 5;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

308. The compound of claim 307, wherein:

d is 4 or 5;

e is 4 or 5; and

f is 1 ;

or a pharmaceutically acceptable salt thereof.

309. The compound of claim 308, wherein m is 2, or a pharmaceutically acceptable salt thereof.

310. The compound of claim 308 wherein m is 3, or a pharmaceutically acceptable salt thereof.

31 1 . The compound of claim 308, wherein m is 4, or a pharmaceutically acceptable salt thereof.

312. The compound of any one of claims 301 -31 1 , wherein

or a pharmaceutically acceptable salt thereof.

313. The compound of claim 312, wherein E is ^OH , or a pharmaceutically acceptable thereof. A compound of formula (XXXII)

(XXXII)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each Y is independently selected from

, -C(0)CH2CH2-, -

CH2CH2NHC(0)CH₂CH2-, and -CH₂-, or is absent;

m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5;

each e is independently from 1 to 15; and

g is from 1 to 15;

or a pharmaceutically acceptable salt thereof.

315. The compound of claim 314, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

316. The compound of claim 315, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

317. The compound of claim 316, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a p armaceutca y accepta e sat t ereo .

318. The compound of claim 317, wherein the combination of Q¹, Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

319. The compound of any one of claims 314-318, wherein Y is -CH2CH2NHC(0)CH₂CH2-, or a pharmaceutically acceptable salt thereof.

320. The compound of any one of claims 314-319, wherein:

m is 2, 3, or 4;

d is an integer from 1 to 5;

e is an integer from 1 to 5;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

321 . The compound of claim 320, wherein:

d is an integer from 1 to 4; and

e is 1 ;

or a pharmaceutically acceptable salt thereof.

322. The compound of claim 321 , wherein d is 2, or a pharmaceutically acceptable salt thereof.

323. The compound of claim 321 , wherein m is 2, or a pharmaceutically acceptable salt thereof.

324. The compound of claim 321 wherein m is 3, or a pharmaceutically acceptable salt thereof.

325. The compound of claim 321 , wherein m is 4, or a pharmaceutically acceptable salt thereof.

326. The compound of any one of claims 314-325, wherein

or a pharmaceutically acceptable salt thereof.

327. The compound of claim 326, wherein E is

, or a pharmaceutically acceptable salt thereof.

A compound of formula (XXXIII):

(XXXIII)

wherein:

each A¹ and A² is an independently selected amino acid;

each E is independently selected from a monosaccharide or an oligosaccharide;

each X is independently selected from

, -C(0)CH₂CH₂C(0)-, -CH2CH2NHC(0)CH₂CH2-, -C(O)-, and -CH₂-, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 alkenyl, optionally substituted C1 -C20 alkynyl, optionally substituted C1 - C15 aryl, optionally substituted C1 -C15 heteroaryl, or is absent;

each Y is independently selected from -CH-, or oxygen ;

Z is -NH-, -NHC(O)-, oxygen, or sulfur;

each m is independently 0, 1 , 2, 3, 4, or 5;

each n is independently 1 , 2, 3, 4, or 5; Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ are each independently selected from the side chain of an amino acid; d is an integer from 1 to 15;

g is an integer from 1 to 15;

e and e' are independently from 1 to 10 and

or a pharmaceutically acceptable salt thereof.

329. The com

(XXXIII-1 )

or a pharmaceutically acceptable salt thereof. 330. The com

(XXXIII-2)

or a pharmaceutically acceptable salt thereof.

331 . The compound of any one of claims 328-330, wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from C1 -C4 alkyl, C1 -C2 hydroxyalkyi, C1 -C5 alkamino, and C6-C35 alkaryl, or a pharmaceutically acceptable salt thereof.

332. The compound of claim 331 , wherein each of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is independently selected from 2-methyl-1 -propyl, 2-propyl, 1 -hydroxyethyl, butyl, benzyl, cyclohexylmethyl, hydroxymethyl, propyl, 2-butyl, methyl, and 2-aminoethyl, or a pharmaceutically acceptable salt thereof.

333. The compound of claim 332, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

334. The compound of claim 333, wherein the combination of Q¹ , Q², Q³, Q⁴, Q⁵ and Q⁶ is selected from one of

or a pharmaceutically acceptable salt thereof.

335. The compound of any one of claims 328-334, wherein:

m is 2, 3, or 4;

d is an integer from 1 to 5;

e is 1 ;

X is -C(0)CH₂CH₂C(0)-;

E is a monosaccharide; and

n is 1 ;

or a pharmaceutically acceptable salt thereof.

336. The compound of claim 335, wherein:

each A¹ and A² is independently selected from 2,4-diaminobutyric acid, 2-aminohexanoic acid, and threonine; and

d is 1 ;

or a pharmaceutically acceptable salt thereof.

337. The compound of claim 336, wherein m is 2, or a pharmaceutically acceptable salt thereof.

338. The compound of claim 336 wherein m is 3, or a pharmaceutically acceptable salt thereof.

339. The compound of claim 336, wherein m is 4, or a pharmaceutically acceptable salt thereof.

340. The compound of any one of claims 328-339, wherein

or a pharmaceutically acceptable salt thereof.

^^'" 'OH

341 . The compound of claim 340, wherein E is ^OH , or a pharmaceutically acceptable salt thereof.

342. The compound of claim 3, wherein the compound is described by formula (XXXIV):

(XXXIV)

each of R², R³, R⁴, R⁵, R⁶, R⁷, R⁸, R⁹, R¹⁰, R'², R'³, R'⁴, R'⁵, R'⁶, R'⁷, R'⁸, R'⁹, and R'¹⁰ is, independently, a lipophilic moiety, a positively charged moiety, a polar moiety, H, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5- C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C2-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; or two R groups on the same or adjacent atoms join to form an optionally substituted 5-8 membered ring;

each of a', b', c', a, b, and c is, independently, 0 or 1 ;

each of N¹ , N², N³, N⁴, N'¹ , N'², N'³, and N'⁴ is a nitrogen atom;

each of C¹ , C², C³, C'¹ , C'², and C'³ is a carbon atom ;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

343. The compound of claim 342, wherein the compound comprises at least one optionally substituted 5- 8 membered ring formed by joining (i) R², R³, and C¹ ; (ii) R³, R⁴, N¹ , and C¹ ; (iii) R⁵, R⁶, and C²; (iv) R⁶, R⁷, N², and C²; (v) R⁸, R⁹, and C³; (vi) R⁹, R¹⁰, N³, and C³; (vii) R'², R'³, and C'¹ ; (viii) R'³, R'⁴, N'¹ , and C'¹ ; (ix) R'⁵, R'⁶, and C'²; (x) R'⁶, R'⁷, N'², and C'²; (xi) R'⁸, R'⁹, and C'³; or (xii) R'⁹, R'¹⁰, N'³, and C'³.

344. The compound of claim 342 or 343, wherein L is described by formula (L-VI):

L^c

L^B-Q-L^A

(L-VI)

G^B2;

G^C2;

G^A1 is a bond attached to Q in formula (L-VI);

G^A2 is a bond attached to A1 or M1 if A1 is absent;

G^B1 is a bond attached to Q in formula (L-VI);

G^B2 is a bond attached to A2 or M2 if A2 is absent;

G^C1 is a bond attached to Q in formula (L-VI);

G^C2 is a bond attached to E;

each of Z^A1 , Z^A2, Z^A3, Z^A4, Z^A5, Z^B1 , Z^B2, Z^B3, Z^B4, Z^B5, Z^C1 , Z^C2, Z^C3, Z^C4 and Z^C5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20 heteroalkylene, optionally substituted C2-C20 alkenylene, optionally substituted C2-C20 heteroalkenylene, optionally substituted C2-C20 alkynylene, optionally substituted C2-C20 heteroalkynylene, optionally substituted C3-C20 cycloalkylene, optionally substituted C3-C20 heterocycloalkylene, optionally substituted C4-C20 cycloalkenylene, optionally substituted C4-C20 heterocycloalkenylene, optionally substituted C8-C20 cycloalkynylene, optionally substituted C8-C20 heterocycloalkynylene, optionally substituted C5-C1 5 arylene, or optionally substituted C2-C15 heteroarylene;

345. The compound of claim 342 or 343, wherein L is described by formula (L-VII):

(L-VII)

G^B2;

G^C2;

G^D2;

G^A1 is a bond attached to Q in formula (L-VII);

G^A2 is a bond attached to A1 or M1 if A1 is absent;

G^B1 is a bond attached to Q in formula (L-VII);

G^B2 is a bond attached to A2 or M2 if A2 is absent;

G^C1 is a bond attached to Q in formula (L-VII);

G^D1 is a bond attached to N in formula (L-VII);

each of Z^A1 Z^A2 Z^A3 Z^M Z^A5 Z^B1 Z^B2 Z^B3 Z^B4 Z^B5 Z^C1 Z^C2 Z^C3 Z^C4 Z^C5 Z^D1 Z^D2 Z^D3 Z^D4 and Z^D5 is, independently, optionally substituted C1 -C20 alkylene, optionally substituted C1 -C20

R' is H, optionally substituted C1 -C20 alkyl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C2-C20 alkenyl, optionally substituted C2-C20 heteroalkenyl, optionally substituted C2-C20 alkynyl, optionally substituted C2-C20 heteroalkynyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C5-C15 aryl, or optionally substituted C2-C15 heteroaryl ;

each of g1 , hi , i1 , j1 , k1 , 11 , ml , n1 , o1 , g2, h2, i2, j2, k2, I2, m2, n2, o2, g3, h3, i3, j3, k3, I3, m3, n3, o3, g4, h4, i4, j4, k4, I4, m4, n4, and o4 is, independently, 0 or 1 ;

346. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXV):

(XXXV)

each of R², R⁶, R⁸, R'², R'⁶, and R'⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyl, optionally substituted C3- C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl; each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

347. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXV):

(XXXV)

each of R², R⁶, R⁸, R'², and R'⁶ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyl, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3- C20 heterocycloalkyi, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

348. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXVI):

(XXXVI) wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

349. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXVII):

(XXXVII)

each of R², R⁶, and R⁸ is, independently, a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4-C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3-C20 heterocycloalkyl, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

350. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXVIII):

(XXXVIII)

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

351 . The compound of any one of claims 342-345, wherein the compound is described by formula (XXXIX):

(XXXIX)

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

352. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXX):

(XXXX)

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

353. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXXI):

(XXXXI)

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

354. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXXI I):

(XXXXI I)

R² is a positively charged moiety, a polar moiety, optionally substituted C1 -C5 alkamino, optionally substituted C1 -C20 alkyl, optionally substituted C3-C20 cycloalkyi, optionally substituted C4- C20 cycloalkenyl, optionally substituted C8-C20 cycloalkynyl, optionally substituted C5-C15 aryl, optionally substituted C1 -C20 heteroalkyi, optionally substituted C3-C20 heterocycloalkyi, optionally substituted C4-C20 heterocycloalkenyl, optionally substituted C8-C20 heterocycloalkynyl, optionally substituted C3-C15 heteroaryl, optionally substituted C6-C35 alkaryl, or optionally substituted C6-C35 heteroalkaryl;

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

355. The compound of any one of claims 342-345, wherein the compound is described by formula (XXXXIII):

(XXXXIII)

each E is, independently, a monosaccharide or oligosaccharide moiety;

n is 1 , 2, 3, or 4,

or a pharmaceutically acceptable salt thereof.

356. The compound of claim any one of claims 342-355, wherein L comprises at least one an optionally substituted C3 cycloalkylene or an optionally substituted C3 heterocycloalkylene.

357. The compound of claim 356, wherein the compound is described by formula (XXXXIV):

(XXXXIV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl; and L comprises an optionally substituted C3 cycloalkylene or an optionally substituted C3 heterocycloalkylene,

or a pharmaceutically acceptable salt thereof.

358. The compound of claim 356, wherein the compound is described by formula (XXXXV):

(XXXXV)

wherein each of R'¹ and R¹ is, independently, optionally substituted benzyl, optionally substituted C2-C15 heteroaryl, optionally substituted C1 -C8 alkyl, or cyclohexylmethyl; and

or a pharmaceutically acceptable salt thereof.

359. The compound of any one of claims 356-358, wherein L is

wherein each q is, independently, an integer from 1 to 1 1 , inclusive.

360. The compound of claim 342-355, wherein L comprises at least one an optionally substituted C5 cycloalkylene or an optionally substituted C5 heterocycloalkylene. The compound of claim 360, wherein the compound is described by formula (XXXXVI):

(XXXXVI)

or a pharmaceutically acceptable salt thereof.

362. The compound of claim 360, wherein the compound is described by formula (XXXXVM):

(XXXXVM)

or a pharmaceutically acceptable salt thereof.

363. The compound of claim 360, wherein the compound is described by formula (XXXXVMI):

(XXXXVMI)

or a pharmaceutically acceptable salt thereof.

364. The compound of claim 360, wherein the compound is described by formula (XXXXIX):

(XXXXIX)

or a pharmaceutically acceptable salt thereof.

65. The compound of any one of claims 360-364, wherein L is

wherein each q is, independently, an integer from 1 to 1 1 , inclusive.

366. The compound of any one of claims 342-355, wherein L comprises at least one an optionally substituted C6 cycloalkylene, at least one an optionally substituted C6 heterocycloalkylene, or at least one an optionally substituted C6 arylene.

367. The compound of claim 366, wherein the compound is described by formula (XXXXX):

(XXXXX)

L comprises an optionally substituted C6 arylene,

or a pharmaceutically acceptable salt thereof.

368. The compound of claim 366 or 367, wherein L is

wherein each q is, independently, an integer from 1 to 1 1 , inclusive.

369. The compound of any one of claims 1 -368, wherein a concentration of the compound, or a pharmaceutically acceptable salt thereof, that activates an immune cell is less than or equal to 10,000 nM.

370. The compound of claim 369, wherein the concentration of the compound, or a pharmaceutically acceptable salt thereof, that activates an immune cell is less than or equal to equal to 1 ,000 nM.

371 . The compound of claim 370, wherein the concentration of the compound, or a pharmaceutically acceptable salt thereof, that activates an immune cell is less than or equal to equal to 100 nM.

372. A pharmaceutical composition comprising a compound of any of claims 1 -368, or a

pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

373. The pharmaceutical composition of claim 372, further comprising an antibacterial agent.

374. The pharmaceutical composition of claim 373, wherein the antibacterial agent is selected from the group consisting of linezolid, tedizolid, posizolid, radezolid, retapamulin, valnemulin, tiamulin, azamulin, lefamulin, plazomicin, amikacin, gentamicin, gamithromycin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, streptomycin, spectinomycin, geldanamycin, herbimycin, rifaximin, loracarbef, ertapenem, doripenem, imipenem/cilastatin, meropenem, cefadroxil, cefazolin, cefalotin, cefalexin, cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepime, ceftaroline fosamil, ceftobiprole, teicoplanin, vancomycin, telavancin, dalbavancin, oritavancin, clindamycin, lincomycin, daptomycin, solithromycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin, aztreonam, furazolidone, nitrofurantoin, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin g, penicillin v, piperacillin, penicillin g, temocillin, ticarcillin, amoxicillin clavulanate,

trimethoprim, prodrugs thereof, and pharmaceutically acceptable salts thereof.

375. The pharmaceutical composition of claim 374, wherein a prodrug of tedizolid is tedizolid phosphate.

376. The pharmaceutical composition of claim 374, wherein the antibacterial agent is tedizolid, azithromycin, meropenem, amikacin, levofloxacin, rifampicin, linezolid, erythromycin, or solithromycin.

377. The pharmaceutical composition of claim 376, wherein the antibacterial agent is tedizolid, azithromycin, meropenem, amikacin, or levofloxacin.

378. A method of protecting against or treating a bacterial infection in a subject, the method comprising administering to the subject a compound of any one of claims 1 -368.

379. The method of claim 378, further comprising administering to the subject an antibacterial agent.

380. A method of protecting against or treating a bacterial infection in a subject, the method comprising administering to the subject (1 ) a compound of any one of claims 1 -368 and (2) an antibacterial agent.

381 . The method of any one of claims 378-380, wherein the bacterial infection is caused by Gram- negative bacteria.

382. The method of any one of claims 378-381 , wherein the bacterial infection is caused by a resistant strain of bacteria.

383. The method of claim 382, wherein the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene, and/or a chromosomal mutation conferring polymyxin resistance.

384. The method of claim 383, wherein the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, or the mcr-3 gene.

385. The method of claim 383, wherein the resistant strain of bacteria possesses a chromosomal mutation conferring polymyxin resistance.

386. The method of any one of claims 382-385, wherein the resistant strain of bacteria is a resistant strain of E. coli.

387. A method of protecting against or treating sepsis in a subject, comprising administering to the subject a compound of any one of claims 1 -368.

388. A method of preventing lipopolysaccharides (LPS) in Gram-negative bacteria from activating an immune system in a subject, comprsing administering to the subject a compound of any one of claims 1 - 368.

389. The method of claim 388, wherein the method prevents LPS from activating a macrophage.

390. The method of claim 388 or 389, wherein the method prevents LPS-induced nitric oxide (NO) production from a macrophage.

391 . The method of any one of claims 388-390, wherein the Gram-negative bacteria is a resistant strain of Gram-negative bacteria.

392. The method of claim 391 , wherein the resistant strain of Gram-negative bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene, and/or a chromosomal mutation conferring polymyxin resistance.

393. The method of claim 392, wherein the resistant strain of Gram-negative bacteria possesses the mcr- 1 gene, the mcr-2 gene, or the mcr-3 gene.

394. The method of claim 392, wherein the resistant strain of Gram-negative bacteria possesses a chromosomal mutation conferring polymyxin resistance.

395. The method of any one of claims 391 -394, wherein the resistant strain of Gram-negative bacteria is a resistant strain of E. coli.

396. The method of any one of claims 387-395, further comprising administering to the subject an antibacterial agent.

397. The method of claim 379, 380, and 396, wherein the compound and the antibacterial agent are administered substantially simultaneously.

398. The method of claim 379, 380, and 396, wherein the compound and the antibacterial agent are administered separately.

399. The method of claim 398, wherein the compound is administered first, followed by administering of the antibacterial agent alone.

400. The method of claim 398, wherein the antibacterial agent is administered first, followed by administering of the compound alone.

401 . The method of claim 379, 380, and 396, wherein the compound and the antibacterial agent are administered substantially simultaneously, followed by administering of the compound or the antibacterial agent alone.

402. The method of claim 379, 380, and 396, wherein the compound or the antibacterial agent is administered alone first, followed by administering of the compound and the antibacterial agent substantially simultaneously.

403. The method of any one of claims 379, 380, and 396-402, wherein administering the compound and the antibacterial agent together lowers the MIC of each of the compound and the antibacterial agent relative to the MIC of each of the compound and the antibacterial agent when each is used alone.

404. The method of any one of claims 378-403, wherein the compound and/or the antibacterial agent is administered intramuscularly, intravenously, intradermally, intraarterially, intraperitoneally, intralesionally, intracranially, intraarticularly, intraprostatically, intrapleural^, intratracheally, intranasally, intravitreally, intravaginally, intrarectally, topically, intratumorally, peritoneally, subcutaneously, subconjunctival, intravesicularlly, mucosally, intrapericardially, intraumbilically, intraocularally, orally, locally, by inhalation, by injection, or by infusion.

405. A method of preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria, comprising contacting the bacteria or a site susceptible to bacterial growth with a compound of any of claims 1 -368.

406. The method of claim 405, further comprising contacting the bacteria or the site susceptible to bacterial growth with an antibacterial agent.

407. A method of preventing, stabilizing, or inhibiting the growth of bacteria, or killing bacteria, comprising contacting the bacteria or a site susceptible to bacterial growth with (1 ) a compound of any of claims 1 -368 and (2) an antibacterial agent.

408. The method of any one of claims 379, 380, 396, 397, and 398, wherein the antibacterial agent is selected from the group consisting of linezolid, tedizolid, posizolid, radezolid, retapamulin, valnemulin, tiamulin, azamulin, lefamulin, plazomicin, amikacin, gentamicin, gamithromycin, kanamycin, neomycin, netilmicin, tobramycin, paromomycin, streptomycin, spectinomycin, geldanamycin, herbimycin, rifaximin, loracarbef, ertapenem, doripenem, imipenem/cilastatin, meropenem, cefadroxil, cefazolin, cefalotin, cefalexin, cefaclor, cefamandole, cefoxitin, cefprozil, cefuroxime, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepime, ceftaroline fosamil, ceftobiprole, teicoplanin, vancomycin, telavancin, dalbavancin, oritavancin, clindamycin, lincomycin, daptomycin, solithromycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin, aztreonam, furazolidone, nitrofurantoin, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin g, penicillin v, piperacillin, penicillin g, temocillin, ticarcillin, amoxicillin clavulanate, ampicillin/sulbactam, piperacillin/tazobactam, ticarcillin/clavulanate, bacitracin, ciprofloxacin, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloxacin, moxifloxacin, nalidixic acid, norfloxacin, ofloxacin, trovafloxacin, grepafloxacin, sparfloxacin, temafloxacin, mafenide, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfadimethoxine, sulfamethizole, sulfamethoxazole, sulfanamide, sulfasalazine, sulfisoxazole, trimethoprim-sulfamethoxazole (tmp-smx),

sulfonamidochrysoidine, eravacycline, demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline, clofazimine, dapsone, capreomycin, cycloserine, ethambutol(bs), ethionamide, isoniazid, pyrazinamide, rifampicin, rifabutin, rifapentine, streptomycin, arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platensimycin, quinupristin/dalfopristin, thiamphenicol, tigecycline, tinidazole, and trimethoprim, prodrugs thereof, and pharmaceutically acceptable salts thereof.

409. The method of claim 408, wherein a prodrug of tedizolid is tedizolid phosphate.

410. The method of claim 408, wherein the antibacterial agent is tedizolid, azithromycin, meropenem, amikacin, levofloxacin, rifampicin, linezolid, erythromycin, or solithromycin.

41 1 . The method of claim 410, wherein the antibacterial agent is tedizolid, azithromycin, meropenem, amikacin, or levofloxacin.

412. The method of any one of claims 405-41 1 , wherein the bacteria are Gram-negative bacteria.

413. The method of any one of claims 405-412, wherein the bacteria are a resistant strain of bacteria.

414. The method of claim 413, wherein the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, the mcr-3 gene, and/or a chromosomal mutation conferring polymyxin resistance.

415. The method of claim 414, wherein the resistant strain of bacteria possesses the mcr- 1 gene, the mcr-2 gene, or the mcr-3 gene.

416. The method of claim 414, wherein the resistant strain of bacteria possesses a chromosomal mutation conferring polymyxin resistance.

417. The method of any one of claims 413-416, wherein the resistant strain of bacteria is a resistant strain of E. coli.

418. A compound selected from any one of compounds 1 -123, or a pharmaceutically acceptable salt thereof.

419. A compound selected from any one of claims 18-47, 71 -124, or 342-368, wherein each of R'¹ and R¹ is, independently, benzyl or CH₂CH(CH₃)2.