EA040967B1 - 1,2,4-OXADIAZOLE AND THIADIASOLE COMPOUNDS AS IMMUNOMODULATORS - Google Patents

Description

This application claims priority from Indian Provisional Application No. 1180/CHE/2015 filed March 10, 2015 and Indian Provisional Application No. 1178/CHE/2015 filed March 10, 2015, the descriptions of which are incorporated herein. document by reference in its entirety.

Technical field

The present invention relates to 1,2,4-oxadiazole and thiadiazole compounds and their derivatives suitable for therapeutic use as immunomodulators. The invention also relates to pharmaceutical compositions containing 1,2,4-oxadiazole and thiadiazole compounds and their derivatives.

State of the art

Programmed cell death protein-1 (PD-1) is a member of the CD28 superfamily that transmits negative signals when interacting with its two ligands PD-L1 or PD-L2. PD-1 and its ligands are widely expressed and exhibit a wider range of immunoregulatory roles in T cell activation and tolerance compared to other members of CD28. PD-1 and its ligands are involved in attenuating infectious immunity and tumor immunity and promoting chronic infection and tumor progression. The biological significance of PD-1 and its ligand indicates the therapeutic potential of manipulation of the PD-1 pathway in various human diseases (HyunTak Jin, et al., Curr Top Microbiol Immunol. (2011), 350:17-37).

T cell activation and dysfunction depends on direct and modulated receptors. Based on their functional output, co-signaling molecules can be divided into co-stimulators and co-inhibitors that positively and negatively regulate the priming, growth, differentiation, and functional maturation of the T cell response (Li Shi, et al., Journal of Hematology & Oncology, 2013 6:74).

Therapeutic antibodies that block the programmed death-1 (PD-1) protein pathway checkpoint prevent T cell downregulation and stimulate anti-cancer immune responses. Several inhibitors of the PD-1 pathway have shown sustained activity across clinical trials (RD Harvey, Clinical Pharmacology & Therapeutics).

Programmed death-1 (PD-1) is a co-receptor that is predominantly expressed by T cells. Binding of PD-1 to its PD-L1 or PD-L2 ligands is vital for the physiological regulation of the immune system. The main functional role of the PD-1 signaling pathway is the inhibition of autoreactive T cells, which serve to protect against autoimmune diseases. Thus, elimination of the PD-1 pathway may lead to the breakdown of immune tolerance, which may ultimately lead to the development of pathogenic autoimmunity. Conversely, tumor cells can sometimes co-opt the PD-1 pathway to escape immune surveillance mechanisms. Thus, blockade of the PD-1 pathway has become an attractive target in cancer therapy. Current approaches include six agents that are either PD-1 or PD-L1 targeted neutralizing antibodies or fusion proteins. More than forty clinical trials are currently underway to better define the role of PD-1 blockade in various tumor types (Ariel Pedoeem et al., Clinical Immunology (2014), 153(1), 145-152).

In international applications Wo 2002086083, Wo 2004004771, Wo 2004056875, Wo 2006121168, Wo 2008156712, Wo 2010077634, Wo 2011066389, Wo 2014055897 and Wo 2014100079 reported PD-1, PD-L1 inhibitory antibodies and/or methods of such antithells. In addition, US patents such as US 8,735,553 and US 8,168,757 report inhibitory PD-1 or PD-L1 antibodies and/or fusion proteins.

In addition, WO 2011161699, WO 2012168944, WO 2013144704 and WO 2013132317 report peptides or peptidomimetic compounds that are capable of suppressing and/or inhibiting the programmed cell death 1 (PD1) signaling pathway.

However, there is a need for more active, better and/or selective immunomodulators of the PD-1 pathway.

The essence of the invention

The present invention provides 1,2,4-oxadiazole and thiadiazole compounds and their pharmaceutically acceptable salts or stereoisomers. These compounds are capable of suppressing and/or inhibiting the programmed cell death-1 (PD1) signaling pathway.

- 1 040967

In one aspect, the present invention provides 1,2,4-oxadiazole and thiadiazole compounds of formula (I)

or a pharmaceutically acceptable salt or stereoisomer thereof;

where ----- is an optional double bond;

X is O or S;

R ₁ and R ₂ independently represent an amino acid side chain or hydrogen, (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ ) alkynyl or cycloalkyl, where (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl and cycloalkyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, carboxylic acid ester, thiocarboxylate, thioacid , -CONR ₇ R ₈ , hydroxy, cycloalkyl, (cycloalkyl)alkyl, aryl, arylalkyl, heterocyclyl, (heterocyclyl)alkyl, heteroaryl, (heteroaryl)alkyl, guanidino, -SH and -S(alkyl); optionally where cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halo, amino, nitro, cyano or alkyl, and optionally where two or three carbon atoms of (C1- _C6 ) alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring);

R ₃ is hydrogen, -CO-[Aaa1]m, [Aaa1]m, [Aaa1]m-CO-[Aaa1]m, -S(O)p-[Aaa1]m, -CONR ₇ R ₈ -COR _c , -SO ₂ Rc, (C 1 -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ ) alkynyl; where (C1-C ₆ )alkyl, (C ₂ C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, -COO-alkyl, carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR7R8, hydroxy, aryl, arylalkyl, cycloalkyl, heterocyclyl, heteroaryl, (cycloalkyl)alkyl, (heterocyclyl)alkyl, (heteroaryl)alkyl, guanidino, -SH and -S(alkyl); optionally where cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl optionally where two or three carbon atoms of (C1- _C6 )alkyl, ( _C2 -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring);

R ₄ and R 5 are independently hydrogen or absent;

R ₆ is hydrogen, alkyl, alkenyl, alkynyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, amino, aminoalkyl, hydroxyalkyl, alkoxyalkyl, acyl, [Aaa2] _w -CO-[Aaa2] _and , [ Aaa2] _n -CO-[Aaa2] _n or -S(O) _p -[Aaa2] _n ;

R ₇ and R ₈ independently represent hydrogen, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl, aryl or heterocyclyl; where (C 1 -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl, aryl and heterocyclyl are optionally substituted with one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, cycloalkyl , heterocyclyl, heteroaryl, aryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl; optionally where two or three carbon atoms of (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring );

alternatively R ₇ and R ₈ together with the nitrogen atom to which they are attached form an optionally substituted 3-7 membered ring containing 0-2 additional heteroatoms independently selected from N, O and S in any stable combination; where the optional Deputy in each case is selected from hydroxyl, -COOH, -COO-alkyl, amide, halogen, amino, nitro and cyano;

[Aaa1] and [Aaa2] are independently at each occurrence an amino acid residue; where the C-terminal carboxyl group of the amino acid residue is a free C-terminal carboxyl group (-COOH) or a modified C-terminal carboxyl group and the N-terminal amino group of the amino acid residue is a free N-terminus (-NH2) or a modified N-terminal amino group ;

R _a is hydrogen or alkyl, alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl or alkoxyalkyl;

Rb is hydrogen or alkyl, alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl or alkoxyalkyl; or R _b and R ₂ together with the atoms to which they are attached may form pyrrolidine or piperidine optionally substituted with one or more groups independently selected from hydroxyl, halogen, amino, cyano and alkyl;

- 2 040967

R _c is (C1-C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl; where the specified (C1-C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl is optionally substituted with one or more substituents selected from carboxylic acid, hydroxyl, alkyl, alkoxy, amino, alkylamino, acylamino, carboxylic acid ester, cycloalkyl, heterocyclyl, heteroaryl , (cycloalkyl)alkyl, (heterocyclyl)alkyl or (heteroaryl)alkyl;

m and n are independently integers selected from 1-3;

p is an integer selected from 1-2;

with the proviso that R1 is not a Ser or Thr side chain when _R2 is an Asp, Asn, Glu or Gln side chain, _R3 is hydrogen, -CO-Ser or -CO-Thr, R6 is hydrogen, alkyl or acyl, Ra and Rb are hydrogen.

In another aspect, the present invention provides a process for the preparation of compounds of formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof.

In a further aspect, the present invention provides pharmaceutical compositions containing a compound of formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, and a process for preparing such compositions.

In yet another aspect, the present invention provides methods for administering a compound of formula (I) or a pharmaceutically acceptable salt or stereoisomer to suppress and/or inhibit the programmed cell death 1 (PD1) signaling pathway. For example, these compounds can be used to treat one or more diseases characterized by impaired or unwanted activity of the PD1 signaling pathway.

Detailed description of the invention

The present invention provides 1,2,4-oxadiazole and thiadiazole compounds and derivatives thereof as therapeutic agents suitable for the treatment of disorders by immunostimulation, including the inhibition of an immunosuppressive signal induced by PD-1, PD-L1 or PD-L2, and therapies, in which these compounds are used.

Each embodiment is presented to illustrate the invention and not to limit the invention. In fact, those skilled in the art will appreciate that various modifications and variations may be made to the compounds, compositions, and methods described herein without departing from the scope or spirit of the invention. For example, features illustrated or described as part of one embodiment may be applied to another embodiment to produce yet another embodiment. Thus, the present invention is intended to include such modifications and variations and their equivalents. Other objects, features and aspects of the present invention are disclosed or obvious from the following detailed description. A person skilled in the art should be clear that the present discussion is a description of exemplary embodiments only and should not be construed as limiting the broader aspects of the present invention.

In some embodiments, the present invention provides compounds of formula (I)

Ra N-------XR _b

Ri (I) or a pharmaceutically acceptable salt or stereoisomer thereof;

where ----- is an optional double bond;

X is O or S;

R ₁ and R ₂ independently represent an amino acid side chain or hydrogen, (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ ) alkynyl or cycloalkyl, where (O ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl and cycloalkyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, carboxylic acid ester, thiocarboxylate, thioacid , -CONR ₇ R ₈ , hydroxy, cycloalkyl, (cycloalkyl)alkyl, aryl, arylalkyl, heterocyclyl, (heterocyclyl)alkyl, heteroaryl, (heteroaryl)alkyl, guanidino, -SH and -S(alkyl); optionally where cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halo, amino, nitro, cyano or alkyl, and optionally where two or three carbon atoms of (C1- _C6 ) alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring);

R ₃ is hydrogen, -CO-[Aaa1] _m , [Aaa1] _m , [Aaa1] _m -CO-[Aaa1] _m , -S(O) _p -[Aaa1] _m ,

- 3 040967

-CONR7R8, -CORc, -SO2R _c , (C1-C6) alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ ) alkynyl; where (C1-C6) alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ ) alkynyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, -COO-alkyl, carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR7R8, hydroxy, aryl, arylalkyl, cycloalkyl, heterocyclyl, heteroaryl, (cycloalkyl)alkyl, (heterocyclyl)alkyl, (heteroaryl)alkyl, guanidino, -SH and -S(alkyl); optionally where cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl optionally where two or three carbon atoms of (C1- _C6 )alkyl, ( _C2 -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring);

R4 and _R5 are independently hydrogen or absent;

R6 is hydrogen, alkyl, alkenyl, alkynyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, amino, aminoalkyl, hydroxyalkyl, alkoxyalkyl, acyl, [Aaa2] _n , -CO-[Aaa2] _n , [ Aaa2] _n -CO-[Aaa2] _n or -S(O) _p -[Aaa2] _n ;

R7 and R8 independently represent hydrogen, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl, aryl or heterocyclyl; where (C 1 -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl, aryl and heterocyclyl are optionally substituted with one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, cycloalkyl , heterocyclyl, heteroaryl, aryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl; optionally where two or three carbon atoms of (C ₁ -C ₆ )alkyl, (C ₂ C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring );

alternatively R7 and _R8 together with the nitrogen atom to which they are attached form an optionally substituted 3-7 membered ring containing 0-2 additional heteroatoms independently selected from N, O and S in any stable combination; where the optional Deputy in each case is selected from hydroxyl, -COOH, -COO-alkyl, amide, halogen, amino, nitro and cyano;

[Aaa1] and [Aaa2] are independently at each occurrence an amino acid residue, where the C-terminal carboxyl group of the amino acid residue is a free C-terminal carboxyl group (-COOH) or a modified C-terminal carboxyl group and the N-terminal amino group of the amino acid residue represents a free N-terminus (-NH2) or a modified N-terminal amino group;

Ra is hydrogen or alkyl, alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl or alkoxyalkyl;

R _b is hydrogen or alkyl, alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl or alkoxyalkyl; or R _b and R2 together with the atoms to which they are attached may form pyrrolidine or piperidine optionally substituted with one or more groups independently selected from hydroxyl, halogen, amino, cyano and alkyl;

Rc is (C1-C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl; where the specified (C1-C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl is optionally substituted with one or more substituents selected from carboxylic acid, hydroxyl, alkyl, alkoxy, amino, alkylamino, acylamino, carboxylic acid ester, cycloalkyl, heterocyclyl, heteroaryl , (cycloalkyl)alkyl, (heterocyclyl)alkyl or (heteroaryl)alkyl;

m and n are independently integers selected from 1-3;

p is an integer selected from 1-2;

with the proviso that R1 is not a Ser or Thr side chain when R2 is an Asp, Asn, Glu or Gln side chain, R ₃ is hydrogen, -CO-Ser or -CO-Thr, R ₆ is hydrogen, alkyl or acyl and Ra and R _b are hydrogen.

In some embodiments of a compound of formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof;

where ----- is an optional double bond;

X is O or S;

R1 and R2 independently represent an amino acid side chain or hydrogen, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl, or cycloalkyl; where (C 1 -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl and cycloalkyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, thiocarboxylate, thioacids, -CONR ₇ R ₈ , hydroxy, cycloalkyl, (cycloalkyl)alkyl, aryl, heterocyclyl, heteroaryl, guanidino, -SH and -S(alkyl); optionally wherein cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl;

R ₃ is hydrogen, -CO-[Aaa1] _m , [Aaa1] _m , [Aaa1] _m -CO-[Aaa1] _m , -S(O) _p -[Aaa1] _m , -CONR7R8, -CORc, - SO ₂ R _c , (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl; where (C1- _C6 )alkyl, (C2-C6)alkenyl and (C2-C6)alkynyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR7R8, hydroxy , cycloalkyl, aryl, heterocyclyl, heteroaryl, guanidino, -SH and -S(alkyl); Not

- 4 040967 necessarily where cycloalkyl, aryl, heterocyclyl and heteroaryl are additionally substituted with one or more substituents, such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl;

R ₄ and R 5 are independently hydrogen or absent;

R6 is hydrogen, alkyl, acyl, [Aaa2] _n , -CO-[Aaa2] _n , [Aaa2] _n -CO-[Aaa2] _n or -S(O)p-[Aaa1] _n ;

R ₇ and R ₈ independently represent hydrogen, (C1-C ₈ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl, aryl or heterocyclyl; where (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl, aryl and heterocyclyl are optionally substituted with one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, cycloalkyl, heterocyclyl, heteroaryl, aryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl; optionally where two or three carbon atoms of (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring );

alternatively R ₇ and R ₈ together with the nitrogen atom to which they are attached form an optionally substituted 3-7 membered ring containing 0-2 additional heteroatoms independently selected from N, O and S in any stable combination; where the optional Deputy in each case is selected from hydroxyl, -COOH, -COO-alkyl, amide, halogen, amino, nitro and cyano;

[Aaa1] and [Aaa2] are each independently selected amino acid residues; where the C-terminal carboxyl group of the amino acid residue is a free C-terminal carboxyl group (-COOH) or a modified C-terminal carboxyl group and the N-terminal amino group of the amino acid residue is a free N-terminus (-NH ₂ ) or a modified N-terminal an amino group;

R _a is hydrogen or alkyl;

R _b is hydrogen or alkyl; or R _b and R2 together with the atoms to which they are attached may form pyrrolidine or piperidine optionally substituted with one or more groups independently selected from hydroxyl, halogen, amino, cyano and alkyl;

Rc is (C ₁ -C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl; where the specified (C1-C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl is optionally substituted with one or more substituents selected from carboxylic acid, hydroxyl, alkyl, alkoxy, amino, alkylamino, acylamino, carboxylic acid ester, cycloalkyl, heterocyclyl, heteroaryl , (cycloalkyl)alkyl, (heterocyclyl)alkyl or (heteroaryl)alkyl;

m and n are independently integers selected from 1-3;

p is an integer selected from 1-2;

provided that R1 is not a Ser or Thr side chain when R2 is an Asp, Asn, Glu or Gln side chain, _R3 is hydrogen, -CO-Ser or -CO-Thr, R6 is hydrogen, alkyl or acyl and R _a and R _b are hydrogen.

In some embodiments, the compounds of the invention are represented by formula (I)

R _a -------XR _b "4 (I) where X is O or S;

each dotted line [----] independently represents an optional double bond;

each Ra and Rb independently represent hydrogen or a substituent such as alkyl, alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl or alkoxyalkyl;

R1 is (C1-C ₆ )alkyl substituted with one or more substituents selected from amino, alkylamino, acylamino, heterocyclyl, heteroaryl, guanidino, (heterocyclyl)alkyl and (heteroaryl)alkyl, where any heterocyclyl or heteroaryl contains at least one nitrogen atom and R1 includes a basic nitrogen atom whose conjugate acid has a pKa greater than 3, preferably greater than 5, and optionally where two or three carbon atoms of (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring);

R2 is (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ )alkynyl substituted with one or more substituents selected from carboxylate, carboxylic acid, carboxylic acid ester, thiocarboxylate, thioacid , amido, amino and heterocyclyl, optionally where two or three carbon atoms of (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbo

- 5 040967 cyclic or heterocyclic ring (such as cyclobutyl or oxirane ring);

R ₃ is hydrogen or -CO-Aaa;

Aaa is an amino acid residue, where said amino acid residue contains a side chain that includes a moiety -OH, -O-acyl, -SH, -NH2 or NH(alkyl);

each of R ₄ and R 5 is independently hydrogen or absent; And

R6 is hydrogen, alkyl, alkenyl, alkynyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, amino, aminoalkyl, hydroxyalkyl, alkoxyalkyl or acyl;

or a pharmaceutically acceptable salt thereof.

According to further embodiments of a compound of formula (I)

or a pharmaceutically acceptable salt or stereoisomer thereof;

where ----- is an optional double bond;

X is O or S;

R1 and R2 independently represent an amino acid side chain or (^-C^ alkyl, (C2-C ₆ )alkenyl, or (C2-C ₆ ) alkynyl; where (C1-C ₆ )alkyl, (C2-C ₆ )alkenyl and (C2- _C6 ) alkynyl is substituted with one or more substituents selected from amino, alkylamino, acylamino, -COO-alkyl, cycloalkyl, heterocyclyl, heteroaryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl; optional where two or three carbon atoms of (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring) ;

R ₃ is hydrogen -CO-[Aaa], -CONR ₇ R ₈ , (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl; where (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl are substituted with one or more substituents selected from amino, alkylamino, acylamino, -COO-alkyl, cycloalkyl, heterocyclyl, heteroaryl , guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl; optionally where two or three carbon atoms of (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring );

R ₄ and R 5 are independently hydrogen or absent;

R6 is hydrogen, alkyl or acyl;

R ₇ and R ₈ independently represent hydrogen, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl; where (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl are substituted with one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, cycloalkyl, heterocyclyl, heteroaryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl; optionally where two or three carbon atoms of (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as cyclobutyl or oxirane ring);

alternatively R ₇ and R ₈ together with the nitrogen atom to which they are attached form an optionally substituted 3-7 membered ring containing 0-2 additional heteroatoms independently selected from N, O and S in any stable combination; where the optional Deputy in each case is selected from hydroxyl, -COOH, -COO-alkyl, amide, halogen, amino, nitro or cyano;

[Aaa] is an amino acid residue;

R _a is hydrogen or alkyl;

R _b is hydrogen or alkyl;

or R _b and R2, together with the atoms to which they are attached, may form pyrrolidine or piperidine optionally substituted with one or more groups independently selected from hydroxyl, halogen, amino, cyano and alkyl; and provided that R1 is not a Ser, Thr, Lys, Arg, or His side chain, when _R2 is an Asp, Asn, Glu, or Gln side chain, _R3 is hydrogen, -CO-Ser, or -COThr, and R _a and R _b are hydrogen.

In certain preferred embodiments of Formula (I), X is O. In certain such embodiments, the ring containing X is an oxadiazole ring.

In some embodiments, R ₁ or R ₂ is an amino acid side chain. Alternatively, R ₁ or R ₂ may be hydrogen.

In some embodiments, R ₁ and R ₂ may independently be

- 6 040967 (C1-Cb) alkyl, (C2-C ₆ )alkenyl, (C ₂ -C ₆ ) alkynyl or cycloalkyl where (C 1-Cb) alkyl, (C ₂ -C ₆ ) alkenyl, (C ₂ -C ₆ ) alkynyl and cycloalkyl are substituted with carboxylic acid, carboxylate, carboxylic acid ester, thiocarboxylate, thio acid, -CONR7R8, hydroxy, cycloalkyl and aryl. In some such embodiments, R1 and R2 may independently be (C1- _C6 )alkyl, ( _C2 - _C6 )alkenyl, or ( _C2 - _C6 )alkynyl substituted with a carboxylic acid ester, thiocarboxylate, thioacid, or cycloalkyl.

In some embodiments, R1 is an amino acid side chain or hydrogen, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ ) alkynyl, or cycloalkyl, where (C 1 -C ₆ )alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl and cycloalkyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, carboxylic acid ester, thiocarboxylate, thioacid, -CONR7R8, hydroxy, cycloalkyl , (cycloalkyl)alkyl, aryl, heterocyclyl, (heterocyclyl)alkyl, heteroaryl, (heteroaryl)alkyl, guanidino, SH and -S(alkyl); optionally where cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halo, amino, nitro, cyano or alkyl, and optionally where two or three carbon atoms of (C1- _C6 ) alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring).

In some embodiments, R1 is (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ )alkynyl; where (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR7R8, hydroxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, guanidino, -SH and -S(alkyl).

In some embodiments, R1 is (C1- _C6 ) alkyl, ( _C2 - _C6 ) alkenyl, or (C2-C6) alkynyl substituted with one or more substituents selected from amino, alkylamino, acylamino, heterocyclyl, heteroaryl, and guanidino , and optionally further substituted with one or more substituents such as alkyl, alkoxy, aralkyl, or aryl.

In some embodiments, R1 is (C1- _C6 ) alkyl, ( _C2 - _C6 ) alkenyl, or ( _C2 - _C6 ) alkynyl substituted with amino, alkylamino, acylamino, heterocyclyl, heteroaryl, guanidino, (heterocyclyl)alkyl ; where (heteroaryl)alkyl, heterocyclyl or heteroaryl contains at least one basic nitrogen atom. Basic nitrogen refers to nitrogen that is protonated and positively charged at pH levels below its pKa value. For example, the pKa of the conjugate acid of the nitrogen-containing compound is >5, preferably >7. In some such embodiments, R ₁ is optionally further substituted with one or more substituents such as alkyl, alkoxy, aralkyl, or aryl.

In some embodiments, R1 is (C1- _C6 ) alkyl, ( _C2 - _C6 ) alkenyl, or (C2-C6) alkynyl substituted with one or more substituents selected from carboxylate, carboxylic acid, carboxylic acid ester, thiocarboxylate , thioacids, -CONR7R8, hydroxy, cycloalkyl, aryl, guanidino, -SH and -S(alkyl). In some such embodiments, R1 is (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ ) alkynyl substituted with one or more substituents selected from a carboxylic acid ester, a thiocarboxylate, thioacid or cycloalkyl.

According to any of the above embodiments, in some embodiments, R1 is (C1- _C4 )alkyl substituted with one or more substituents selected from amino, heteroaryl, or guanidino. In some embodiments, R ₁ is -(CH ₂ )imidazolyl, (CH2) ₃ NHC(=N)-NH2 or -(CH ₂ ) ₄ NH ₂ .

In some embodiments, R2 is an amino acid side chain or hydrogen, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl, or cycloalkyl; where (C1- _C6 )alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl and cycloalkyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, carboxylic acid ester, thiocarboxylate , thioacids, -CONR7R8, hydroxy, cycloalkyl, (cycloalkyl)alkyl, aryl, heterocyclyl, (heterocyclyl)alkyl, heteroaryl, (heteroaryl)alkyl, guanidino, SH and -S(alkyl); optionally where cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl, and optionally where two or three carbon atoms of (C ₁ -C ₆ )alkyl, ( C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring);

In some embodiments, R2 is (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ )alkynyl; where (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR7R8, hydroxy, cycloalkyl, aryl, heterocyclyl, (heterocyclyl)alkyl, heteroaryl, (heteroaryl)alkyl, guanidino, -SH and -S(alkyl). In some such embodiments, R2 is (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ ) alkynyl, substituted

- 7 040967 with one or more substituents selected from alkylamino, acylamino, cycloalkyl and (heterocyclyl)alkyl.

In some embodiments, R2 is (C1- _C6 )alkyl substituted with one or more substituents selected from carboxylate, carboxylic acid, carboxylic acid ester, thiocarboxylate, thioacid, amido, amino, and heterocyclyl, and optionally further substituted with one or more substituents such as alkyl, alkoxy, aralkyl or aryl. In some such embodiments, R2 further optionally contains one or more double bonds or triple bonds. In some embodiments, R2 is (C ₃ -C ₈ )cycloalkyl substituted with one or more substituents selected from carboxylate, carboxylic acid, thiocarboxylate, thioacid, amido, ester, amino, and heterocyclyl, and further optionally substituted with one or more additional substituents such as alkyl, alkoxy, aralkyl or aryl.

According to any of the above embodiments, in some embodiments, R2 is (C1- _C4 )alkyl substituted with one or more substituents selected from carboxylate, carboxylic acid, and amido. In some embodiments, R ₂ is -(CH ₂ )CoOH, -(CH ₂ ) ₂ COOH, -(CH2)CONH2, or -(CH2)2CONH2. In certain such embodiments, R2 is -(CH2)2C(O)NH2, -CH2C(O)NH2, -( _CH2 ) _2C (O)NH(alkyl), or _-CH2C (O)NH(alkyl ).

In some embodiments, R2 is (C1- _C6 ) alkyl, ( _C2 - _C6 ) alkenyl, or ( _C2 - _C6 ) alkynyl substituted with one or more substituents selected from amino, alkylamino, acylamino, hydroxy, cycloalkyl , aryl, (heterocyclyl)alkyl, heteroaryl, (heteroaryl)alkyl, guanidino, -SH and -S(alkyl). In some such embodiments, R2 is (C ₁ -C 6 )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ ) alkynyl substituted with one or more substituents selected from alkylamino, acylamino, cycloalkyl, and (heterocyclyl )alkyl.

In some embodiments, R ₃ is hydrogen, -CO-[Aaa1] _m , [Aaa1] _m , [Aaa1] _m -CO-[Aaa1] _m or -S(O) _p -[Aaa1] _m .

In some embodiments, R ₃ is -CO-Aaa1 and the side chain of Aaa1 contains a (C1-C ₄ )alkyl group optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR ₇ R ₈ , hydroxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, guanidino, -SH, -S(alkyl); optionally wherein cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl.

Alternatively, R ₃ may be -CO-[Aaa1] _m , where m is greater than 1. In other embodiments, R ₃ may be [Aaa1] _m , [Aaa1] _m -CO-[Aaa1] _m or -S( O) _p -[Aaa1] _m , where m is an integer from 1 to 3.

In additional embodiments, the Aaa1 side chain contains a (C ₁ -C ₄ )alkyl group substituted with one or more substituents selected from amino, acylamino, carboxylic acid, -CONR7R8, hydroxy, cycloalkyl, aryl, heteroaryl, guanidino, -SH, and - S(alkyl); where R7 and R8 independently represent hydrogen, alkyl, aryl or heterocyclyl.

In further alternative embodiments, R ₃ may be (C 1 -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ ) alkynyl, substituted with carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR7R8, hydroxy, -SH and -S(alkyl). In particular, R ₃ is CONR7R8; where R7 and R8 independently represent hydrogen, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl; where (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl are substituted with one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, cycloalkyl, heterocyclyl, heteroaryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl.

In additional alternative embodiments, R3 may be -CONR7R8; where R7 and R8 independently represent hydrogen, (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl; where (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl and (C ₂ -C ₆ )alkynyl; optionally where two or three carbon atoms of (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring ).

In some embodiments, R ₃ is -CORc or -SO2R where R _c is (C1-C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl; where the specified (C ₁ -C ₆ )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl is optionally substituted with one or more substituents selected from carboxylic acid, hydroxyl, alkyl, alkoxy, amino, alkylamino, acylamino, carboxylic acid ester, cycloalkyl, heterocyclyl, heteroaryl, (cycloalkyl)alkyl, (heterocyclyl)alkyl or (heteroaryl)alkyl.

Alternatively, R3 may be -CORc or -SO2Rc, where Rc is (C1- _C6 )alkyl, cycloalkyl, aryl, heterocyclyl or heteroaryl; where the specified (C ₁ -C ₆ )alkyl, aryl, heterocyclyl or heteroaryl is optionally substituted with one or more substituents selected from carboxylic acid, hydroxyl, alkyl, amino or acylamino.

In additional alternative implementations, R ₃ may be

- 8 040967 (C1-C ₆ )alkyl, (C2-C ₆ )alkenyl or (C2-C ₆ )alkynyl, be substituted by carboxylic acid, carboxylate, thiocarboxylate, thioacid, -CONR7R8, hydroxy, -SH and -S(alkyl ).

In some embodiments, R6 is hydrogen, alkyl, [Aaa2] _n or -CO-[Aaa2] _n . R6 may be -CO-[Aaa2] _n . Alternatively, R6 may be H.

In some embodiments, R6 is -CO-Aaa2 and the Aaa2 side chain contains a (C1- _C4 ) alkyl group optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, thiocarboxylate, thioacid, - CONR7R8, hydroxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, guanidino, -SH, -S(alkyl); optionally wherein cycloalkyl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl.

In additional embodiments, the Aaa2 side chain contains a (C ₁ -C ₄ )alkyl group substituted with one or more substituents selected from amino, acylamino, carboxylic acid, -CONR7R8, hydroxy, cycloalkyl, aryl, heteroaryl, guanidino, -SH, and - S(alkyl); where R ₇ and R ₈ independently represent hydrogen or alkyl.

In some embodiments, Aaa1 or Aaa2 is an amino acid residue, wherein said amino acid residue contains a side chain that includes an OH, -O-acyl, -SH, -NH2, or NH(alkyl) moiety.

In some embodiments, R ₇ is (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ ) alkynyl substituted with one or more substituents selected from halogen, hydroxyl, amino, nitro, cyano, cycloalkyl, heterocyclyl, heteroaryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl.

In some embodiments, R ₈ is (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ ) alkynyl substituted with one or more substituents selected from halogen, hydroxyl, amino, nitro , cyano, cycloalkyl, heterocyclyl, heteroaryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl and (heteroaryl)alkyl.

In some embodiments, R _a is alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl, or alkoxyalkyl.

In some embodiments, R ₁ is an amino acid side chain or hydrogen, (C 1 -C ₆ )alkyl, (C ₂ -C ₆ ) alkenyl, (C ₂ -C ₆ ) alkynyl, or cycloalkyl; where (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ )alkynyl and cycloalkyl are optionally substituted with one or more substituents selected from amino, alkylamino, acylamino, carboxylic acid, carboxylate, ester carboxylic acid, thiocarboxylate, thioacid, -CONR ₇ R ₈ , hydroxy, cycloalkyl, (cycloalkyl)alkyl, aryl, heterocyclyl, (heterocyclyl)alkyl, heteroaryl, (heteroaryl)alkyl, guanidino, SH, and -S(alkyl); optionally where cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl, and optionally where two or three carbon atoms of (C ₁ -C6) alkyl, (C ₂ -C ₆ )alkenyl or (C ₂ -C ₆ )alkynyl form part of a 3-7 membered carbocyclic or heterocyclic ring (such as a cyclobutyl or oxirane ring); And

Ra is hydrogen or alkyl, alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl or alkoxyalkyl.

In some embodiments, R _b is alkenyl, alkynyl, acyl, aralkyl, aryl, heteroaralkyl, heteroaryl, cycloalkyl, (cycloalkyl)alkyl, aminoalkyl, hydroxyalkyl, or alkoxyalkyl. Alternatively, in some embodiments, R _b and R2, together with the atoms to which they are attached, form a pyrrolidine or piperidine ring, optionally substituted with one or more groups independently selected from hydroxyl, halogen, amino, cyano, and alkyl.

In some embodiments, the present invention provides compounds of formula (IA)

or a pharmaceutically acceptable salt or stereoisomer thereof;

where R1, R2, R ₃ , R ₆ , R _a and R _b are as defined in formula (I).

In some embodiments of a compound of formula (I) or formula (IA), R _b is H.

In additional embodiments of a compound of formula (I) or formula (IA), R ₃ is -CO-[Aaa1] _m .

- 9 040967

For example, the compounds of the present invention may have the structure of formula (IB)

or a pharmaceutically acceptable salt or stereoisomer thereof;

where R1, R2, R5, Ra, [Aaa1] and m are as defined in formula (I).

In some embodiments, the present invention provides formulas (IC)

or a pharmaceutically acceptable salt or stereoisomer thereof;

where R1, R2, [Aaa1] and m are as defined in formula (I).

In some embodiments, the present invention provides formulas (ID)

or a pharmaceutically acceptable salt or stereoisomer thereof;

where R1, R2, R3, R _a , R _b , [Aaa2] and n are as defined in formula (I).

In some embodiments, the present invention provides formulas (IE) compound compound compound

covalent bond between the carbonyl carbon and the amino group of an amino acid residue. In preferred embodiments, the amino acid is an alpha amino acid and the amino acid residue Aaa is connected to the carbonyl group of CO via a covalent bond between the carbonyl carbon and the alpha amino group of the amino acid residue.

According to any of the above implementations, there implementations of X is O.

In accordance with any of the above implementation options there implementation of formula (I), (IA) or (ID) R 3 represents hydrogen.

According to one of the above embodiments, there implementations of formula (I), (IA) or (ID) R3 is -CO-Aaa.

In accordance with any of the above embodiments according to according to according to according to according to some embodiments of some embodiments of formula (I), (IA), (IB), (IC), (ID) or (IE) R1 is alkyl substituted with amino or heteroaryl. Preferably R1 is -(CH ₂ ) ₄ NH ₂ .

According to any of the above embodiments, according to some embodiments, R ₁ is an amino acid side chain.

According to any of the above embodiments, according to some embodiments, R ₂ is an amino acid side chain.

- 10 040967

According to any of the above embodiments, in some embodiments, R1 is a Lys, Tyr, Gln, Ser, Ala, Glu, Leu, Asp, or His side chain.

In accordance with any of the above implementation options according to some variants of implementation of R ₁ represents the side chain of Lys, Tyr or Gln.

According to any of the above embodiments, according to some embodiments, R ₁ is a side chain of Tyr.

In accordance with any of the above implementation options according to some variants of implementation of R ₁ represents the side chain of Lys.

According to any of the above embodiments, according to some embodiments, R1 is a Gln side chain.

In alternative embodiments, R1 is not a Ser or Thr side chain; those. R1 is not -CH2OH or -CH( _CH3 )OH. In additional alternative embodiments, R1 is not a Ser or Thr side chain that is alkylated or acylated. For example, in some embodiments, Ri is not -CH2OCH3, -CH2OAc, -CH(CH ₃ )OCH ₃ , or -CH(CH ₃ )OAc. In additional embodiments, R1 is not a Ser or Thr side chain, which are optionally alkylated or acylated, i. R1 is not -CH2OH, -CH( _CH3 )OH, -CH2OCH3, -CH2OAc, -CH( _CH3 ) _OCH3 or -CH( _CH3 )OAc.

According to any of the above embodiments, according to some embodiments of formula (I), (IA), (IB), (IC), (ID) or (IE), R2 is alkyl substituted with amido. In some embodiments, R2 is -(CH2) ₂ C(O)NH2 or -CH2C(O)NH2. Preferably R2 is -CH2C(O)NH2.

According to any of the above embodiments, in some embodiments, R2 is a Gly, Gln, Glu, Ser, Asn, Asp, Ala, or Ile side chain.

In accordance with any of the above implementation options according to some variants of implementation of R ₂ represents the side chain of Gln, Glu, Asn, Asp or Ile.

According to any of the above embodiments, according to some embodiments, R ₂ is an Ile side chain.

According to any of the above embodiments, according to some embodiments, R ₂ is an Ala side chain.

According to any of the above embodiments, according to some embodiments, R2 is the Asn side chain.

In alternative embodiments, R2 is not an Asn, Asp, Gln, or Glu side chain; those. R2 is not -CH2C(O)NH2 _{, -CH2C} (O)OH, _-CH2CH2C (O) _NH2 or -CH2CH2C(O)OH.

According to any of the above embodiments, in some embodiments, Rb and R2, together with the atoms to which they are attached, may form a pyrrolidine optionally substituted with hydroxyl.

According to any of the above embodiments, according to some embodiments, m is 1.

According to any of the above embodiments, according to some embodiments, [Aaa1] contains a side chain that includes a -OH moiety.

According to any of the above embodiments, in some embodiments, [Aaa1] is Ala, Thr, Ile, Glu, Lys, Asp, Tyr, Gln, Ser, or Phe.

According to any of the above embodiments, according to some embodiments, [Aaa1] is Ser, Thr, Tyr, Glu, Ala, or Ile.

According to any of the above embodiments, according to some embodiments, [Aaa1] is Tyr.

According to any of the above embodiments, according to some embodiments, [Aaa1] is Glu.

According to any of the above implementation options, according to some implementation options, [Aaa1] is Ala.

In accordance with any of the above implementation options according to some implementation options [Aaa1] is Thr.

In accordance with any of the above embodiments, according to some embodiments, [Aaa1] is He.

In alternative embodiments, R ₃ is -CO-[Aaa1] and Aaa1 is not a Thr or Ser amino acid residue.

In additional alternative embodiments, R3 is not H or -CO-[Aaa1].

According to any of the above embodiments, according to some embodiments, n is 1.

- 11 040967

According to any of the above embodiments, in some embodiments, [Aaa2] is Ala, Thr, Ile, Glu, Lys, Asp, Tyr, Gln, Ser, or Phe.

According to any of the above embodiments, according to some embodiments, [Aaa2] is Ser, Thr, Tyr, Glu, Ala, or Ile.

According to any of the above embodiments, according to some embodiments, p is 2.

According to any of the above embodiments, in some embodiments, one, more or all of the amino acid residues are D amino acid residues.

According to any of the above embodiments, in some embodiments, one, more than one, or all of the amino acid residues are L amino acid residues.

In some embodiments, the present invention provides a compound, or a pharmaceutically acceptable salt or stereoisomer thereof, selected from

Connection No. Structure 1 1 ^? About Hz ^N th J 2 °γ ^0Η νη ₂ 4 οΑ _ο γΟΗ ^\ ⁿ ^'n ^A nV ^h Λ—Λ ^{Η Η} δ 3 ΝΗ ₂ I ^Η % οΑ _Ο Α Αό ^{Η Η} δ 4 νη ₂ °Υ ^{οη Η} % οΑ o g ΑΑ ^{Η Η} δ

- 12 040967

- 13 040967

14 But. YJL " \ AH 1 - o γ η^^Λν V" Ν—O ^{N N} O 15 nh ₂ XT O \/°n । o z o γ ι!Ι—A ^{n n} A 16 BUT. L / „ \YUN O X O Of Η,ΝΑχΟΧχ L—A ^{n n} A 17 BUT. „ \/OH O \u003d ° f ι! Ι - A ^{n n} A 18 0 ^but OH ₂ A 0 . H ₂ N ιί—A ^{N N} A 19 ^s^OH XX but ^ ₀ XX η ₂ ν ^Λ Υ ^ν Υ ^Λ ν ^Λ ν ^Λ υ ^οη L—A ⁿ A 20 t 1 gt X O? X A .on H ₂ N Of NN Of II 1 H II N O O 21 BUT.^^ XX. /\o γ ^0Η ΗϊνΧυΧνΛΟ ¹ ν—A ⁿ A 22 o ^on ^x ^on nLDHyT™ Ν--O O

- 14 040967

23 OH he o^^ Z\ o ¢^0 M ⁿ S 24 °γ ^ΟΗ on A A o M ⁿ S 25 xXj O %-OH Η,Ν ^Af 'Ch^'' NN L—A ⁿ O 26 ^ho ^a ry ^OH Λ—A ^HH δ 27 i^N^° H BUT. to \/OH 1 \u003d ⁰ T H ₂ N 'Af NN 7< ^OH N-O ^HH O 28 nh ₂ °V ^nh ' ^H % oA 0 / η^Α-^ν^^νΑτ^ |!]_i H ⁿ D 29 ,nh ₂ νη ₂ A \ ^OH ° | Ao JL X. ,OH h ₂ n At Ar nn Ar II 1 η η II N --- O O thirty Η ₂ Ν^ΝΗ < ^NH o ^nL o f ^OH i'—i ^HH S

- 15 040967

- 16 040967

39 NH ₂ ⁿ % "A o AD iLU'L / L A ¹ -A ^HH A 40 nh ₂ \ / xL \ /OH y^ o y o y^ /C .N. Ox JL L. /OH Η ₂ Ν tG Ν Ν II 1 n n II Ν --- O O 41 ΝΗ ₂ ^ΗΝ \χ>χ /X . *X/ ^OH L O x o H ₂ N% ^N ^y A—A ^{n n} A 42 BUT. 1st T ² S / S Eh X ^ON । o o y^ /C ,N. Ox /C xC /OH Η ₂ Ν ν Ν II 1 n n II Ν --- O O 43 IT. ^ΗΝ \Α \ /OH I = ⁰ 1Г xL /Ν. L. JL /C /OH H ₂ N hl Ν Ν TG II 1 Η Η II Ν ---Ο 0 44 ^Η0 \ \ \ /OH 1 ⁰ y^ xC χΝ^ /C xC A /OH h ₂ n Y ^NN IT II 1 η η II N---0 0 45 r^NH ₂ NH ₂ X q4. o XX h ₂ nOvVOv ⁰ ^ J—A ^{n H} A 46 s ^x nh ₂ oh X 0^0 o /U _o tAWW J—A ^HH S 47 H.O. H.O. „ ,OH L—A ^HH A

- 17 040967

48 he he he oA o/'- O h,h\Y' ^A V ^H S—4 ^{n n} S 49 L ^NH2 A oA o γ ^0Η ^γΥΑΛ™ L—O ^{n n} S 50 AA νη ₂ <A oA ₀ uen AyYAnA AA ^{n n} o 51 νη ₂ NO. xA \ yun 1 ° A o DI xA χΝ. Ah xA χΆχ ^Ν \χ^χ ^Ο \-AAi Ar'0j 52 L x- L x- \ yun AC V o D. χΝ. L. JL D. jun h ₂ n AA nn II 1 HH II N---0 0 53 i? X Z O An and Y IZ * O O o \u003d (o o ^x I 54 nh ₂ oA o'V ^OH η^Ατ'-Α'νΑνΑτ ⁰ Λ—A ^HH S 55 nh ₂ oh °γ ^ΟH o^^ o A h ⁰ H ₂ NNN f ^N A^OH i ¹ —i ^HH S a ^/ ^NH 2 56 nh ₂ nh ₂ nh _2oA . oA О Ao η,νΑχΝ^Λ,,Α,^χΑοη AA ^HH S

- 18 040967

57 Η,Ν % Ch^on ? 9 _g °n VnA J Η O ² NO Η N \-N // ^H g _; Oh 58 nh ₂ ^HS \ /L \ .OH % ⁰ = Ο xL .N. JI L. .OH H ₂ NYNN > r II 1 η η II N --- O O 59 -AhV' 4 < 0 Y ^0H H^LY^Y ⁰ ιϋ-ο ⁿ s 60 H.O. AO, λ H ₂ N^ ^N ^ _n AQ^OH N—Ο Η H 61 N—. O <1 A OH I 1 ^H * ^N % ? ( _u ..A ,N/-.AA___on H2N Τι ίΓ NN ri N—0 ^HH O 62 jQ χ,ΟΗ ^h * ⁿ N—0 H 63 i ^ NH ₂ OH L -___OH O ^ > Ο γ W / OH H ₂ N η NNY 11 ¹ η h 11 N--O O 64 H oh ^nh ₂ < ^N y ^NH o^\ 4 0 r ^NH2 η^υΥ'Λ'Υ'π L-i Η hn

- 19 040967

- 20 040967

73 NH, □A _o > H ₂ l/ N-° ^HH O 74 Q ο 'ΑγΑγ ¹ h ₂ n ^Z n-° ^hh o 75 HHO. L _l \/OH A - Ο γ AALA Y—O ^HH O 76 ^ho a k o γ ^ΟΗ AA ^A V ^H N—0 ^HH 0 77 ^ΗΟ ~Ί \A ^{H 7} ? ΊΓ ^h2n ^ ulLmAgon NO Η H 78 ^HN ^° o^nh ₂ NO. Г J η ₂ ν\ύα a JCn^am ANN >f '^YUN ^N ~O η Η Η Ξ ^^OH 79 nh ₂ ⁿ % A o v ^OH II I η h // \ N--0 Nn oh 80 nh ₂ ^H °. A _{0 V} OH L/'VA Ain H ₂ N \\ / NN^if ² N-0 ^H H II 0

- 21 040967

81 °^.X ^H 2 nh ₂ y HO. J > ° 3 ? fH? H ₂ N Ίί^ ΊΓ NN ^V OH II 1 η η II z N--O X. ΌΗ 82 I nh ₂ A °A 9 Η Η 9 n^^y^^y^on ^N -° ⁰ ^OH 83 _NH2HO . \.OH | ° + o X^ ^n. A. JL X. .on H ₂ N if nn if 1 II η η II О---N О 84 x° ZT cX' A ^z . z? I y ^z / z o £ 85 o h ₂ n^A o HO-, .. ? V\X Λ /X xO / \\ 1 η ° H ₂ N nO 86 o H ₂ N^X o HO-λ .. W λ / XN % / \\ AH ° H ₂ N n-° 87 0 H ₂ N^_ 0 I ΗΟ-Λ Μ Λ X YZ^f N % h ₂ n no 88 o H ₂ N^_ o I \ NO-L .. X _ς Α/ \ Xs xo V / XN % / v 1 η o h ₂ n _n -o 89 0 H ₂ N^4 O P ^Η θ~Ά m L / \\ IH O H ₂ N n-°

- 22 040967

90 o ⁿ % H ₂ gA o T _o M but o 91 0 η ₂ ν^4 o O Η ₂ Ν^ _Ν - ^H 92 nh ₂ O^H o ^HO ~\ L L \/ N '<X..YUN / \\ / LI \ / H ₂ N Ν'θ HN—/ 93 nh ₂ 0*4 0 OH ^but l JU- \_/um y ''' H ₂ N n-° ^H nh ₂ 94 nh ₂ 0*4 oh but—. μ l A /\\ I LI I H ₂ N nO 95 0 NgM^Ch ^but l aL-a. /—k 1 n Il ^N H ₂ N _N -0 \ 96 nh ₂ 0*4 О no-, _m 1 1 k 1 s ι/\ ^χ П ^{Н Н} Η2Ν ΝΟ 97 νη ₂ NO. „ \ / OH h o a o y U / W > h UX / OH Η ₂ Ν η N II NX and 1 η 0 h 11 N O O 98 nh ₂ ^h % A 0 ^g h^'V^nInA/'h M ^H0H !

-23 040967

99 ι^^ΝΗ ₂ OH L _o x^OH ² C non J 100 Pnh?Y ^{NH2 H} N o v ^oH h ₂ n\Vn1nV ^h A ^h ° ^h π 101 H ₂ N^NH r ^NH o AL o y™ ² M ^hoh g 102 H /\Z 0^ ^NHO . L ~ \/OH A -° T ² C non 1 103 p _NH ?Y ^NH2 Ί - ° Г° ^н HzN-V^nIn-V™ i ¹ —A h°h Π 104 nh ₂ \ \/OH Λ oA 0 y X .OH h ₂ n TN II NT Il । η ο η II N O 0 105 _N O^OH \ n ^ ^0H L—A ^HH S 106 0 ''^''N'''''! NH ₂ ⁿ L _n \/ ^oh O | O -_ O y ^H Ν—O ^{H H} O

- 24 040967

107 / I IZ o Z=< 1 ^z ° >° IZ y--< o=( ^z o 1? I 108 h ₂ n^nh r ^NH 0 YES. o hAvAy ί—A ^{n H} A 109 HHO. -N. /NH < || < nh ₂ < γ to about > ^NH2 xL /Ύ /A JL /K / OH H ₂ N AA NN Al II 1 η η II N --- O O 110 o^,nh ₂ o^.nh ₂ i T JLJ A A o A^ .N. Y\ .OH H ₂ N A< NN Ar II 1 η η II N---О 0 111 Ν-η O ο _Ύ °Η η=ν\'Άν ^Λ νΥ ^0Η ί—A ^{Η Η} J 112 1? I Z O z=< 1 ^z ° >° / IZ / (. o=( o ( ° ¹ 3 FROM nh ₂ ^H % A ο γ ^{ΟΗ Η} ₂ ^Ν ^τΓ ^Ν ν^ ^Ν ^ ^Ν ^ιΓ°\-θΗ ² IA ^{n n} YES 114 ΥΊ? \u003d ⁰ ϊ / A ^ n. A. JL / H / He Η ₂ Ν Αι Ν Ν At II 1 n n II Ν ---Ο Ο

- 25 040967

115 Η ₂ Ν^^ΝΗ HhL O Y OH Y. O O ^H * ^N ϊ YA η I i ^he Ν---° ° ΥΊ 116 H ₂ N_O ^Η 2^\χ^θ TT Г T 1 Y ° /N. χΥ _OH H ₂ N II NN II 1 η η II N---0 0 117 AY ^^Y ° xY χΝ. A .OH H ₂ NNN II 1 η η II N---0 0 118 1 Y A oA ο γ ^θΗ "LUAD-™ L—A ^HH S 119 H ₂ N^,NH .NH O^_NH ₂ ) ) χΟΗ f Of Η ₂ νΑχ/ ^Ν ^χ^ _ν ^Υ _ν Α^ΟΗ II 1 η ο η II N---O O 120 Ν-, O \Υ ,, _kl Υ \х ^on th ) η ₂ ν·Υ ο Ύ hAWA° ^{H 2} Щ ΗΟΗ ; 121 Ϊ ^Η 2 ^Ν Υ Υ ^ΗΟ = θ g J-A ^non A 122 Λ ^ΝΗ2 οΑ ο ύ°η ² Αο ^ΗΟΗ ί, 123 ΥΑ ^ΝΗ2 Ύ <Α _ξ 0 γΟΗ η ₂ ν ^Υτ ν Α ν 1< ^ΟΗ Μ ^ηοη S " 124 1 Υ ^{Η2 S} _\ \/ ^ΟΗ A O A o y A^ γ A A /On Η

or a pharmaceutically acceptable salt thereof or a stereoisomer thereof.

In some embodiments, the compounds of the invention may be prodrugs of compounds of formula (I) wherein, for example, the hydroxyl in the parent compound is present as an ester or carbonate, or the carboxylic acid present in the parent compound is present as an ester. In a further embodiment, the prodrug is metabolized to the active parent compound in vivo (eg, the ester is hydrolyzed to form the corresponding hydroxyl or carboxylic acid).

In some embodiments, the compounds of the present invention may also contain, in non-natural proportions, isotopes of atoms of one or more elements that are

- 26 040967 put such connections. For example, the present invention also extends to isotopically labeled embodiments of the present invention which are identical to those specified herein, but in which one or more atoms of the compound are replaced by atoms having atomic masses or mass numbers other than the predominant atomic masses or mass numbers that are normally occur naturally for given atoms. All present isotopes of any particular atom or element are within the group of compounds of the present invention and their uses. Typical isotopes that can be introduced into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine and iodine, such as ² H (D), ³ H, ^P C, ¹³ C, ¹⁴ C, ¹³ N, ¹⁵ N, ¹⁵ O, ¹⁷ O, ¹⁸ O, ³⁵ S, ¹⁸ F, ³⁶ Cl, ¹²³ I and ¹²⁵ I. , which are described below in the schemes and/or in the examples provided in this document, by replacing the reagent without an isotope label with an isotopically labeled reagent.

pharmaceutical compositions.

In some embodiments, the present invention provides a pharmaceutical composition comprising a compound described herein, optionally in admixture with a pharmaceutically acceptable carrier or diluent.

The present invention also provides methods for preparing formulations of the disclosed compounds for pharmaceutical administration.

The compositions and methods of the present invention may be used to treat an individual in need thereof. In some embodiments, the individual is a mammal, such as a human or a non-human mammal. When administered to an animal such as a human, the composition or compound is preferably administered as a pharmaceutical composition containing, for example, a compound of the present invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or buffered physiological saline, or other solvents or excipients such as glycols, glycerin, oils such as olive oil, or injectable organic compounds. ethers. In a preferred embodiment, when such pharmaceutical compositions are for human administration, in particular for invasive routes of administration (i.e., routes such as injection or implantation that bypass transport or diffusion across the epithelial barrier), the aqueous solution is pyrogen-free or non-pyrogenic. essentially non-pyrogenic. Excipients may be selected, for example, to effect a sustained release of the agent or selectively affect one or more types of cells, tissues or organs. The pharmaceutical composition may be present in unit dosage form such as tablet, capsule (including coated particle capsule and gelatin capsule), granule, reconstitution lyophile, powder, solution, syrup, suppository, injection, etc. The composition may also be present in a transdermal delivery system such as, for example, a skin patch. The composition may also be present in a solution suitable for topical administration, such as eye drops.

A pharmaceutically acceptable carrier may contain physiologically acceptable agents which act, for example, to stabilize, increase solubility or increase absorption of a compound, such as a compound of the present invention. Such physiologically acceptable substances include, for example, carbohydrates such as glucose, sucrose or dextrans, antioxidants such as ascorbic acid or glutathione, chelating agents, small molecular weight proteins, or other stabilizers or adjuvants. The choice of a pharmaceutically acceptable carrier comprising a physiologically acceptable agent depends, for example, on the route of administration of the composition. The formulation of the pharmaceutical composition may be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) may also be a liposome or other polymeric matrix, which may contain, for example, the compound of the present invention introduced into it. For example, liposomes that include phospholipids or other lipids are non-toxic, physiologically acceptable and metabolizable carriers that are relatively easy to manufacture and administer.

The expression "pharmaceutically acceptable" as used herein means those compounds, materials, compositions and/or dosage forms that are medically suitable for use in contact with human and animal tissues without causing undue toxicity, irritation, allergic reaction, or other problems or complications, in accordance with a reasonable benefit-risk ratio.

The expression pharmaceutically acceptable carrier as used herein means a pharmaceutically acceptable material, composition, or excipient, such as a liquid or solid excipient, diluent, excipient, solvent, or encapsulating material. Each carrier must be acceptable in the sense of being compatible with the other ingredients of the composition and not injurious to the patient. Some examples of materials that can serve in

- 27 040967 as pharmaceutically acceptable carriers include (1) sugars such as lactose, glucose and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols such as propylene glycol; (11) polyhydric alcohols such as glycerol, sorbitol, mannitol and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic solution; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

The pharmaceutical composition (preparation) can be administered to a patient by any of various routes of administration, including, for example, oral (for example, liquid dosage forms, which are aqueous or non-aqueous solutions or suspensions, tablets, capsules (including capsules with coated particles and gelatin capsules), boluses , powders, granules, pastes for application to the tongue); absorption (through the mucous membrane of the oral cavity, for example, sublingual); anal, rectal or vaginal (including, for example, a pessary, cream or foam); parenteral (including intramuscular, intravenous, subcutaneous or intrathecal, for example, in the form of a sterile solution or suspension); nasal; intraperitoneal; subcutaneous; transdermal (eg, in the form of a patch applied to the skin); and topical (including, for example, a cream, ointment or aerosol for application to the skin or eye drops). The compound may also be contained in a composition for inhalation. In some embodiments, the compound may simply be dissolved or suspended in sterile water. A detailed description of appropriate routes of administration and compositions suitable for this purpose can be found, for example, in US patent No. 6110973, 5763493, 5731000, 5541231, 5427798, 5358970 and 4172896, as well as in the patents cited therein.

The formulations may conveniently be prepared in unit dosage forms and may be prepared by any of the methods well known in the pharmaceutical art. The amount of active ingredient with which the carrier material may be combined to form a unit dosage form will vary depending on the patient to be treated, the particular route of administration. The amount of active ingredient with which the carrier material can be combined to form a unit dosage form will generally be that amount of the compound that produces a therapeutic effect. Typically, on a 100% basis, this amount will be from about 1% to about 99%, preferably from about 5% to about 70%, most preferably from about 10% to about 30%.

Methods for preparing said formulations or compositions comprise the step of combining an active compound, such as a compound of the invention, a carrier, and optionally one or more accessory ingredients. In general, formulations are prepared by uniformly and intimately combining the compound of the invention with liquid carriers and/or finely divided solid carriers and, if necessary, subsequent shaping of the product.

Formulations of the present invention that are suitable for oral administration may be present in such forms as capsules (including coated particle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored base, typically sucrose and gum acacia or tragacanth), lyophile, powders, granules, or solution or suspension in an aqueous or non-aqueous liquid, or oil-in-water or water-in-oil liquid emulsion, or elixir or syrup, or lozenge (using an inert base such as gelatin and glycerin or sucrose and acacia), and/or mouthwash, etc., wherein each formulation contains a predetermined amount of a compound of the present invention as an active ingredient. The compositions or compounds may also be administered in the form of a bolus, electuary or paste.

To prepare solid dosage forms for oral administration (capsules (including coated particle capsules and gelatin capsules), tablets, pills, dragees, powders, granules, etc.), one or more pharmaceutically acceptable carriers, such as sodium citrate or dibasic calcium phosphate, and/or any of the following: (1) fillers or bulk fillers such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia; (3) humectants such as glycerin; (4) leavening agents such as agar-agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retaining agents such as paraffin; (6) absorption accelerators such as quaternary ammonium compounds; (7) wetting agents such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents such as kaolin and bentonite clay; (9) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; (10) complex

- 28 040967 forming agents such as modified and unmodified cyclodextrins; and (11) colorants. In the case of capsules (including coated particle and gelatin capsules), tablets and pills, the pharmaceutical compositions may also include buffering agents. Solid compositions of a similar type may also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

The tablet may be made by compression or moulding, optionally with one or more accessory ingredients. Compressed tablets can be prepared using a binder (eg, gelatin or hydroxypropyl methylcellulose), a lubricant, an inert diluent, a preservative, a disintegrant (eg, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), a surfactant, or a dispersing agent. Molded tablets can be made by molding a mixture of a powdered compound wetted with an inert liquid diluent in a suitable machine.

Tablets and other solid dosage forms of pharmaceutical compositions such as dragees, capsules (including coated particle capsules and gelatin capsules), pills and granules may optionally be provided with notches or coatings and shells such as enteric coatings and other coatings well known in the art. in the field of pharmaceutical manufacturing. They can also be formulated to provide a slow or controlled release of the active ingredient contained therein using, for example, hydroxypropyl methylcellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be sterilized, for example, by filtration through a bacteria-retaining filter, or by administering sterilizing agents in the form of sterile solid compositions which may be dissolved in sterile water or some other injectable sterile medium immediately prior to use. Said compositions may also optionally contain opacifying agents and may be formulated to release one or more active ingredients exclusively or preferentially in a certain part of the gastrointestinal tract, optionally in a delayed manner. Examples of matrix materials that can be used include polymeric materials and waxes. The active ingredient may also be present in microencapsulated form, if appropriate, including one or more of the excipients described above.

Liquid dosage forms suitable for oral administration include pharmaceutically acceptable emulsions, reconstitution lyophiles, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and their derivatives, solubilizing agents, and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (particularly cottonseed, peanut, corn, corn germ oil, olive, castor, and sesame oils), glycerin, tetrahydrofuryl alcohol, polyethylene glycols, and fatty acid esters, and sorbitol, as well as mixtures thereof.

In addition to inert diluents, oral compositions may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweeteners, flavors, colors, flavors and preservatives.

In addition to active compounds, suspensions may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, tragacanth, and mixtures thereof.

Pharmaceutical compositions for rectal, vaginal or urethral administration may be suppositories which may be prepared by mixing one or more active compounds with one or more suitable non-irritating excipients or carriers which contain, for example, cocoa butter, polyethylene glycol, suppository wax or salicylate and which are solid at room temperature but liquid at body temperature and thus will melt in the rectum or vagina and release the active compound.

Formulations of pharmaceutical compositions for oral administration may be a mouthwash or an oral aerosol or oral ointment.

Alternatively, or in addition, the compositions may be designed to be delivered via a catheter, stent, rod, or other intraluminal device. Delivery through such devices may be particularly suitable for use in delivery to the bladder, urethra, ureter, rectum or intestines.

Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or aerosol dosage forms containing such

- 29 040967 existing media, which are known in the art.

Dosage forms for topical or transdermal use include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers or propellants that may be required.

In addition to the active compound, ointments, pastes, creams and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide or their mixtures.

In addition to the active compound, powders and sprays may contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

Transdermal patches have the added benefit of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be prepared by dissolving or dispersing the active compound in an appropriate medium. Absorption enhancers can also be used to increase the flux of a compound through the skin. The rate of such flow can be controlled by using a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

Ophthalmic dosage forms, ophthalmic ointments, powders, solutions, and the like are also included within the scope of the present invention. Exemplary ophthalmic formulations are described in US Pat. If necessary, liquid ophthalmic compositions have properties similar to those of lacrimal fluid, tissue fluid or vitreous moisture, or are compatible with such fluids. The preferred route of administration is topical administration (eg, topical administration such as eye drops, or administration by implantation).

The expressions parenteral administration and parenterally administered as used herein means methods of administration that are not enteral and topical administration and are usually injections, including, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intravertebral and intrasternal injections and infusions.

Pharmaceutical compositions suitable for parenteral administration include one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions or sterile powders which can be reconstituted into sterile injectable solutions or dispersions. immediately before use, which may contain antioxidants, buffer substances, bacteriostats, solutions of which make the dosage form isotonic with respect to the blood of a given recipient, or suspending agents or thickeners.

Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of the present invention include water, ethanol, polyhydric alcohols (such as glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents such as, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic substances such as sugars, sodium chloride, and the like in the compositions. In addition, delayed absorption of an injectable pharmaceutical form can be provided by the inclusion of agents that delay absorption, such as aluminum monostearate and gelatin.

In some cases, in order to prolong the effect of the drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be done by using a liquid suspension of crystalline or amorphous material having low water solubility. The rate of absorption of the drug then depends on its rate of dissolution, which in turn may depend on the size of the crystals and the crystalline form. Alternatively, delayed parenteral absorption

- 30 040967 this dosage form is carried out by dissolving or suspending the drug in an oily carrier medium.

Suitable injectable depot forms are prepared by forming microencapsulated matrices of the desired compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer and the nature of the particular polymer used, the rate of drug release can be controlled. Examples of other biodegradable polymers include polyorthoesters and polyanhydrides. Suitable injectable depot formulations are also prepared by placing the drug in liposomes or microemulsions that are compatible with body tissues.

For use in the methods of the present invention, the active compounds may be present in pure form or in the form of a pharmaceutical composition containing, for example, from 0.1 to 99.5% (preferably from 0.5 to 90%) of the active ingredient in combination with a pharmaceutically acceptable carrier.

Administration methods may also be provided by rechargeable or biodegradable devices. A variety of slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinaceous biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including biodegradable and non-degradable polymers, can be used to form implants for sustained release of a compound at a specific target site.

Actual dosage levels of active ingredients in pharmaceutical compositions can be varied so as to obtain the active ingredient in an amount that will be effective to achieve the desired therapeutic response for a particular patient, composition and form of administration, without toxic effects on the patient.

The selected dosage level will depend on a variety of factors, including the activity of the particular compound used or combination of compounds or the corresponding esters, salts or amides, the route of administration, the time of administration, the rate of release of one or more of the specific compounds used, the duration of treatment, other drugs, compounds and /or materials used in combination with one or more specific compounds used, age, sex, condition, general and medical history of the patient to be treated, as well as other factors well known to specialists in the field of medicine.

A physician or veterinarian skilled in the art can readily determine and prescribe a therapeutically effective amount of the desired pharmaceutical composition. For example, a physician or veterinarian may set initial doses of a pharmaceutical composition or compound at a level less than that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. The term "therapeutically effective amount" means a concentration of a compound that is sufficient to produce the desired therapeutic effect. As a rule, it is understood that the effective amount of the compound will vary depending on the weight, sex, age and history of the subject. Other factors that affect the effective amount may include, but are not limited to, the severity of the patient's condition, the disorder being treated, the stability of the compound, and, if necessary, another type of therapeutic agent being taken with the compound of the present invention. A higher total dose can be delivered by repeated administration of the agent. Methods for determining efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996), Harrison's Principles of Internal Medicine 13 ed., 1814-1882, which is incorporated herein by reference).

In general, a suitable daily dose of the active compound used in the compositions and methods of the present invention will be that amount of the compound which is the minimum dose effective to provide a therapeutic effect. Typically, such an effective dose will depend on the factors described above.

If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally in unit dosage forms. In some embodiments of the present invention, the active compound may be administered two or three times a day. In preferred embodiments, the active compound may be administered once a day.

The patient receiving said treatment is any animal in need thereof, including primates, in particular humans, and other mammals such as horses, cattle, pigs and sheep; as well as poultry and domestic animals, in general.

Wetting agents, emulsifiers and lubricating agents such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavoring and flavoring agents, preservatives and antioxidants may also be present in the compositions.

- 31 040967

Examples of pharmaceutically acceptable antioxidants include (1) water-soluble antioxidants such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

Methods of treatment.

The programmed cell death protein 1 (PD-1) pathway is involved in a number of diseases and conditions and is known to regulate a variety of immune responses. Numerous studies have been conducted to activate the immune response by influencing the PD-1 pathway to thereby provide treatment for certain conditions such as various types of cancer. As such, studies show that blockade of the PD-1 pathway, e.g., by inhibiting the immunosuppressive signal induced by PD-1, PD-LI, or PD-L2, results in antitumor activity in various types of cancer, including lung, breast, colon intestines, kidneys, bladder, thyroid, prostate, osteosarcoma and Hodgkin's lymphoma.

In addition, PD-1 activity is also associated with autoimmune conditions such as lupus erythematosus, juvenile idiopathic arthritis, and allergic encephalomyelitis.

In some embodiments, the present invention provides uses of a compound of the present invention for the manufacture of a medicament, for example, for the treatment of cancer.

In some embodiments, the present invention provides methods for treating cancer, wherein said methods comprise administering to a subject in need thereof a therapeutically effective amount of a compound of the present invention.

In some embodiments, the present invention provides methods for inhibiting tumor cell growth and/or metastasis by administering to a subject in need thereof a therapeutically effective amount of the compounds of the present invention.

In some embodiments, the present invention provides methods for inhibiting tumor cell growth and/or metastasis by administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I).

In some embodiments, the present invention provides methods for treating cancer by administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I).

Typical tumor cells include cancer cells such as, but not limited to, melanoma, kidney cancer, prostate cancer, breast cancer, colon cancer and lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, anal cancer, stomach cancer, testicular cancer, fallopian tube carcinoma, endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, non-Hodgkin's lymphoma, cancer esophagus, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal gland cancer, soft tissue sarcoma, urethral cancer, penile cancer, chronic or acute leukemias, including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, childhood solid tumors, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer, renal pelvis carcinoma ci, central nervous system (CNS) neoplasm, non-small cell lung cancer (NSCLC), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brainstem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers, including asbestos-induced cancers and combinations of these cancers.

In some embodiments, the present invention provides methods for treating cancer, wherein the cancer is selected from lung cancer, breast cancer, colon cancer, kidney cancer, bladder cancer, thyroid cancer, prostate cancer, osteosarcoma, and Hodgkin's lymphoma.

In some embodiments, the present invention provides methods for treating a bacterial, viral, or fungal infection or immunological condition by administering a therapeutically effective amount of a compound of formula (I) or pharmaceutically acceptable salts and corresponding stereoisomers to a subject in need thereof.

In some embodiments, the present invention provides uses of a compound of the present invention for the manufacture of a medicament for the treatment of a bacterial, viral, and fungal infection, as well as methods of administering a therapeutically effective amount of a compound of the present invention for the treatment of a bacterial, viral, or fungal infection.

In some embodiments, the present invention provides applications with

- 32 040967 units of formula (I) for the preparation of a drug for the treatment of a bacterial, viral and fungal infection, as well as methods for administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt and stereoisomer for the treatment of a bacterial, viral or fungal infection.

In other embodiments, the present invention provides a method of treating an infection by blocking the PD-1 pathway, such as inhibiting an immunosuppressive signal induced by PD-1, PD-L1 or PD-L2, wherein said method comprises administering to a subject in need thereof a therapeutically effective amount of a compound according to the present invention.

In some embodiments, the present invention provides uses of a compound of the present invention to inhibit the PD-1 pathway (eg, PD-1, PD-L1, or PD-L2).

In some embodiments, the present invention provides methods for treating an infectious disease in a subject, comprising administering a therapeutically effective amount of a compound of the present invention to treat the infectious disease.

In some embodiments, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt and stereoisomer thereof, for use as a medicament.

In some embodiments, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt and stereoisomer thereof, for use in the treatment of cancer.

In some embodiments, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt and stereoisomer thereof, for use in the treatment of lung cancer, breast cancer, colon cancer, kidney cancer, bladder cancer, thyroid cancer, prostate cancer, osteosarcoma and Hodgkin's lymphoma.

In some embodiments, the present invention provides a compound of formula (I), or a pharmaceutically acceptable salt and stereoisomer thereof, for use in the treatment of a bacterial, viral, or fungal infection or immunological condition.

Typical infectious diseases include, but are not limited to, HIV, influenza, herpes, giardiasis, malaria, leishmania, pathogenic infections that cause hepatitis virus (A, B, and C), herpes virus (e.g., VZV, HSV-I, HAV -6, HSV-II and CMV, Epstein-Barr virus), adenovirus, influenza virus, flaviviruses, echovirus, rhinovirus, coxsackievirus, coronovirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus , Human T-lymphotropic virus, dengue fever virus, papillomavirus, mollusk virus, poliovirus, rabies virus, John Cunningham virus and arbovirus encephalitis virus, pathogenic infections that cause chlamydia bacteria, rickettsia bacteria, mycobacteria, staphylococci, streptococci, pneumococci, meningococci and gonococci, Klebsiella, Proteus, Serratia, Pseudomonas, Escherichia coli, Legionella, diphtheria, salmonella, bacilli, cholera, tetanus, botulism, anthrax, plague, leptospirosis and disease Lyme disease, pathogenic infections caused by fungi of the genera Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, niger, etc.), Genus Mucorales (mucor, absidia, rhizophus) , Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis and Histoplasma capsulatum, and pathogenic infections caused by the parasites Entamoeba histolytica, Balantidium coli, Naegleriafowleri, Acanthamoeba sp., Giardia vivia, Cryptosporidium sp., Pneumocystis carinri, Plasmodium tiax , Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondii, Nippostrongylus brasiliensis.

The compounds of the present invention may be used as single drugs (monotherapy) or together with one or more other agents (combination therapy). The compounds may be used in pure form, or preferably in a pharmaceutical composition in which one or more pharmaceutically acceptable materials are admixed with the compound.

The pharmaceutical composition may be administered by oral or inhalation routes or by parenteral administration. For example, the compositions may be administered orally, by intravenous infusion, topically, intraperitoneally, intravesically, or intrathecally. Examples of parenteral administration include, but are not limited to, intra-articular (into the joints), intravenous, intramuscular, intradermal, intraperitoneal, and subcutaneous routes. Suitable liquid compositions may be aqueous or non-aqueous isotonic sterile injectable solutions and may contain antioxidants, buffering agents, bacteriostats, and soluble agents that make the dosage form isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions, which may contain suspending agents, solubilizers, thickeners, stabilizers and preservatives. Oral administration, parenteral administration, subcutaneous administration and intravenous administration are the preferred routes of administration.

The dosage of the compounds according to the present invention varies with age,

- 33 040967 weight or symptoms of the patient, as well as the activity or therapeutic efficacy of the compound, dosage regimen and/or time of treatment. In general, suitable routes of administration may, for example, include oral, eye drops, rectal, transmucosal, topical or intestinal administration; parenteral delivery, including intramuscular, subcutaneous, bone marrow injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal or intraocular injections. The compounds according to the invention may be administered in amounts of 0.5 or 1 to 500 mg, 1 or 2 g, depending on the dosage regimen. Doses can be administered once a week, every three days, every two days, once a day, twice a day, three times a day, or more frequently. In alternative embodiments for certain adult subjects, the compound may be administered continuously via intravenous injection for a period of time prescribed by the physician. Since dosage is affected by various conditions, in some cases an amount less than or greater than the prescribed dosage may be administered. The physician can easily determine the appropriate dosage for the patient undergoing therapeutic treatment.

Compounds of the present invention may be administered in combination with one or more other drugs to (1) complement and/or enhance the effect of a compound of the present invention, (2) modulate pharmacodynamics, improve absorption, or reduce the dose of a compound of the present invention, and/or (3) reduce or alleviate the side effects of the compound of the present invention. As used herein, the term combined administration means any form of administration of two or more different therapeutic compounds in such a way that the second compound is administered while the previously administered therapeutic compound is still effective in the body (e.g., the two compounds simultaneously produce their effect on the patient, which may include the synergistic effects of the two compounds). For example, various therapeutic compounds may be administered in a single dosage form or in separate dosage forms, including simultaneously or sequentially. In some embodiments, the various therapeutic compounds may be administered within 1, 12, 24, 36, 48, 72 hours or weeks of each other. Thus, an individual who receives such treatment may experience the combined effect of various therapeutic compounds. The respective compounds may be administered in the same or different ways and in the same or different ways.

The dosage of the other drug may be that used in clinical practice, or it may be a reduced dosage that is effective when administered in combination with a compound of the present invention. The ratio of the compound of the present invention to the other drug may vary depending on the age and weight of the subject to be treated, the route of administration, the time of administration, the disease to be treated, the symptoms, and combinations thereof. For example, another drug may be used in an amount of 0.01 to 100 parts by weight. per 1 wt.h. compounds according to the present invention.

Combination therapy can be used to treat any of the diseases discussed herein. For example, in the methods of the present invention for the treatment of cancer, the compound of the present invention may be used through existing chemotherapy in conjunction with the use of a single pharmaceutical composition or a combination of different pharmaceutical compositions. Examples of chemotherapeutic agents include an alkylating agent, a nitrosocarbamide agent, an antimetabolite, an anticancer antibiotic, a plant alkaloid, a topoisomerase inhibitor, a hormonal drug, a hormone antagonist, an aromatase inhibitor, a P-glycoprotein inhibitor, a platinum complex derivative, other immunotherapeutic drugs, and other anticancer drugs. . In addition, the compound of the invention can be co-administered with a cancer adjuvant such as a leukopenia (neutropenia) drug, a thrombocytopenia drug, an antiemetic, and a cancer pain reliever, simultaneously or in the form of mixtures. Chemotherapeutic agents that can be co-administered with the compounds of the invention include the following: aminoglutethimide, amsacrine, anastrozole, asparaginase, bcg, bicalutamide, bleomycin, bortezomib, buserelin, busulfan, campothecin, capecytarabine, carboplatin, carfilzomib, carmustine, chlorambucil, chloroquine, cisplatin , cladribine, clodronate, colchicine, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, demethoxyviridine, dexamethasone, dichloroacetate, dienestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, estradiol, estramustine, etoposide, everolimus, fludrobine, exemestane, fludrobine, exemestane, , fluorouracil, fluoxymesterone, flutamide, gemcytarabine, genistein, goserelin, hydroxyurea, idarubicin, ifosfamide, imatinib, interferon, irinotecan, ironotecan, lenalidomide, letrozole, leucovorin, leuprolide, levanisol, lomustine, lonidamine, meclorethamine, medroxytophalogestrogesterone, meroximestrogestrone, meroximestrogestrone , mesna, metformin, methotrex t, mitomycin, mitotane, mitoxantrone, nilutamide, nocodazole, octreotide, oxaliplatin,

- 34 040967 paclitaxel, pamidronate, pentostatin, perifosine, plicamycin, pomalidomide, porfimer, procarbazine, ratlytrexed, rituskimab, sorafenib, streptozocin, sunitinib, suramin, tamoxifen, temozolomide, temsirolimus, teniproside, testosterone, thalidomide, thioguanine, thiotepa, titanocene dichloride, topotecan, trastuzumab, tretinion, vinblastine, vincristine, vindesine and vinorelbine.

In some embodiments, a compound of the present invention may be administered in conjunction with non-chemical treatments for cancer. According to an additional implementation variant, the compound according to the present invention can be entered in conjunction with radiation therapy. In a further embodiment, a compound of the present invention may be administered in conjunction with surgery, thermal ablation, focused ultrasound therapy, cryotherapy, or any combination of these treatments.

In some embodiments, various compounds of the present invention may be co-administered with one or more other compounds of the present invention. In addition, such combinations may be administered in conjunction with other therapeutic agents, such as other agents suitable for the treatment of cancer, immunological or neurological diseases, such as those listed above. In some embodiments, the co-administration of one or more additional chemotherapeutic agents with a compound of the present invention provides a synergistic effect. In some embodiments, the co-administration of one or more additional chemotherapeutic agents provides an additive effect.

The compound of the present invention can be used with one or more other immunomodulators and/or potentiators when used together in the same pharmaceutical composition or through a combination of different pharmaceutical compositions. Suitable immunomodulators include a variety of cytokines, vaccines, and adjuvants. Typical cytokines, vaccines, and adjuvants that stimulate immune responses include GM-CSF, M-CSF, G-CSF, interferon-α, β or γ, IL-1, IL-2, IL-3, IL-12, Poly (I:C) and C _P G.

In some embodiments, potentiating agents include cyclophosphamide and cyclophosphamide analogs, anti-TGFe and imatinib (Glivec), a mitosis inhibitor such as paclitaxel, sunitinib (Sutent) or other angiogenic agents, an aromatase inhibitor such as letrozole, an A2a-adenosine receptor antagonist ( A2AR), an angiogenesis inhibitor, anthracyclines, oxaliplatin, doxorubicin, TLR4 antagonists and IL-18 antagonists.

Definitions and abbreviations.

Unless otherwise specified, all technical and scientific terms used in this document have the same meanings, and the meanings of such terms are independent in each application and represent the meanings that are usually understood by a person skilled in the art to which this document pertains. Regardless, and except for the definition of other conditions, the following definitions apply throughout the text of the description and claims. Chemical names, common names, and chemical structures can be used interchangeably to describe the same structure. If a chemical compound is described using both the chemical structure and the chemical name, and there is a mismatch between the structure and the name, then the structure takes precedence. These definitions apply whether the term is used alone or in combination with other terms, unless otherwise specified. Therefore, the definition of alkyl applies to alkyl as well as to the alkyl moieties of hydroxyalkyl, haloalkyl, -O-alkyl, etc.

The term compounds according to the present invention includes compounds of formula (I), their pharmaceutically acceptable salts and the corresponding stereoisomers.

The term acyl is known in the art and refers to the group represented by the general formula hydrocarbylC(O)-, preferably alkylC(O)-.

The term acylamino means an amino group substituted with an acyl.

The term alkoxy means an alkyl group, preferably a lower alkyl group, having an oxygen atom attached to it. Representative alkoxy groups include methoxy, ethoxy, propoxy, t-butoxy, and the like.

The term alkenyl, as used herein, means an aliphatic group containing at least one double bond, and includes unsubstituted alkenyls and substituted alkenyls, where the latter means alkenyl moieties having substituents that replace the hydrogen on one or more carbon atoms of the alkenyl group. Such substituents may be present on one or more carbon atoms, which may or may not be included in one or more double bonds. In addition, such substituents include all substituents provided for alkyl groups as discussed above, except where stability is prohibitive. For example, substitution of alkenyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

An alkyl group or alkane is a non-aromatic hydrocarbon with a direct or

- 35 040967 branched chain, which is fully saturated. Typically, a straight or branched alkyl group contains from 1 to about 20 carbon atoms, preferably from 1 to about 10, unless otherwise specified. Examples of straight or branched chain alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, t-butyl, pentyl, hexyl, heptyl, and octyl. A ^-C^ straight chain or branched alkyl group is also referred to as a lower alkyl group. The alkyl group may optionally be substituted at one or more positions, as far as valency permits. Such optional substituents include, for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic or heteroaromatic moieties, -CF ₃ , -CN, and the like.

The term alkylamino as used herein means an amino group substituted with at least one alkyl group.

The term alkylthio as used herein means a thiol group substituted with an alkyl group and may be represented by the general formula alkyl S-.

The term alkynyl as used herein means an aliphatic group containing at least one triple bond and includes unsubstituted alkynyls and substituted alkynyls, where the latter means alkynyl moieties having substituents that replace hydrogen atoms on one or more carbon atoms of the alkynyl group. Such substituents may be present on one or more carbon atoms that are included or not included in one or more triple bonds. In addition, such substituents include all substituents provided for alkyl groups as discussed above, except where stability is prohibitive. For example, substitution of alkynyl groups with one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

The term amide or amido as used herein means the group O

Ry where R ^x and R ^y are each independently hydrogen or a hydrocarbyl group, or both R ^x and R ^y taken together with the N atom to which they are attached form a heterocycle having 4 to 8 atoms in the ring structure.

The terms in amine and amino are recognized in the art and refer to both unsubstituted and substituted amines and their salts, for example, a fragment that can be represented

where each R ^x is independently hydrogen or a hydrocarbyl group, or two R ^x taken together with the N atom to which they are attached form a heterocycle having 4 to 8 atoms in the ring structure.

The term aminoalkyl, as used herein, means an alkyl group substituted with an amino group.

The terms aralkyl and arylalkyl, as used herein, mean an alkyl group substituted with an aryl group.

The term aryl as used herein means substituted or unsubstituted monocyclic aromatic groups in which each ring atom is a carbon atom. Preferably the ring is a 5-7 membered ring, preferably a 6 membered ring. The term aryl also includes polycyclic ring systems having two or more cyclic rings, in which two or more carbon atoms are shared by two adjacent rings, where at least one of the rings is aromatic, for example, other cyclic rings can be cycloalkyls, cycloalkenyls , cycloalkynyls, aryls, heteroaryls and/or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

A cycloalkyl group is a cyclic hydrocarbon that is fully saturated. Cycloalkyl includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group contains from 3 to about 10 carbon atoms, more typically from 3 to 8 carbon atoms, unless otherwise specified. The second bicyclic cycloalkyl ring may be selected from saturated, unsaturated and aromatic rings. Cycloalkyl includes bicyclic molecules in which one, two or three or more atoms are shared between two rings. The term fused cycloalkyl means a bicyclic cycloalkyl in which each ring

- 36 040967 shares two atoms with another ring. The second fused bicyclic cycloalkyl ring may be selected from saturated, unsaturated and aromatic rings. A cycloalkenyl group is a cyclic hydrocarbon containing one or more double bonds. The cycloalkyl group may be substituted at one or more positions, as far as valency permits, and contain any of the optional substituents described herein. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

The term cyano means a -CN group.

The term carboxy or carboxylic acid, as used herein, means a group represented by the formula -CO2H.

The term carboxylate means a group represented by the formula -(CO2) ^- .

The term ester as used herein means a -C(O)OR ^x group, where R ^x is a hydrocarbyl group.

The term guanidino as used herein means the -NH-C(=NH)-NH2 group.

The terms halo and halogen as used herein mean halogen and include chlorine, fluorine, bromine and iodine.

The term haloalkyl as used herein means an alkyl group substituted with a halogen atom.

As used herein, the term carbocycle, carbocyclic, or carbocyclyl means any stable 3-, 4-, 5-, 6-, or 7-membered monocyclic or bicyclic or 7-, 8-, 9-, 10-, 11-, 12-, or 13 -membered bicyclic or tricyclic hydrocarbon ring, any of which may be saturated, partially unsaturated, unsaturated or aromatic. Examples of carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cycloheptenyl, cycloheptyl, cycloheptenyl, adamantanyl, cyclooctyl, cyclooctenyl, cyclooctadienyl, [3.3.0]bicyclooctane, [4.3.0]bicyclononane, [ 4.4.0]bicyclodecane, [2.2.2]bicyclooctane, fluorenyl, phenyl, naphthyl, indanyl, adamantyl, anthracenyl and tetrahydronaphthyl (tetralin). As presented above, bridged rings are also included in the definition of a carbocycle (eg [2.2.2]bicyclooctane). Preferred carbocycles, unless otherwise specified, are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl and indanyl. When the term carbocycle or carbocyclyl is used, it includes the term aryl. Bridged rings are formed when one or more carbon atoms are bonded to two non-adjacent carbon atoms. Preferred bridges consist of one or two carbon atoms. It should be noted that the bridge always converts a monocyclic ring into a tricyclic ring. When the ring is bridged, the substituents specified for the ring may also be present on the bridge.

The terms hetaralkyl and heteroaralkyl as used herein mean an alkyl group substituted with a hetaryl group.

The term heteroalkyl as used herein means a saturated or unsaturated chain of carbon atoms and at least one heteroatom, where no two heteroatoms are adjacent.

The terms heteroaryl and hetaryl mean substituted or unsubstituted aromatic monocyclic structures, preferably 5 to 7 membered rings, more preferably 5 to 6 membered rings, in which the ring structures comprise at least one heteroatom, preferably one to four heteroatoms, preferably one or two heteroatom. The terms heteroaryl and hetaryl also include polycyclic ring systems having two or more cyclic rings in which two or more carbon atoms are shared by two adjacent rings, where at least one of the rings is heteroaromatic, for example, the other cyclic rings may be cycloalkyls , cycloalkenyls, cycloalkynyls, aryls, heteroaryls and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, indole, 1,3,4-oxadiazole, pyrimidine, and the like. The heteroaryl group may be substituted at one or more positions, as far as valency permits, and contain any of the optional substituents described herein.

The term heteroatom as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen and sulfur.

The terms heterocyclyl, heterocycle and heterocyclic mean substituted or unsubstituted non-aromatic ring structures, preferably 3 to 10 membered rings, more preferably 3 to 7 membered rings, in which the ring structures comprise at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms heterocyclyl and heterocyclic also include polycyclic ring systems having two or more cyclic rings in which two or more carbon atoms are shared by two adjacent rings, where at least one of the rings is heterocyclic, for example, the other cyclic rings may be cycloalkyls , cycloalkenyls, cycloalkynes

- 37 040967 ly, aryl, heteroaryl and/or heterocyclyl. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, 2,3-dihydrobenzo[b][1,4]dioxin, lactones, lactams, and the like. Heterocyclyl groups may be optionally substituted as far as valence permits.

The term heterocyclylalkyl or (heterocyclyl)alkyl as used herein means an alkyl group substituted with a heterocyclic group.

The term hydroxyalkyl as used herein means an alkyl group substituted with a hydroxyl group.

As used herein, the term hydroxy or hydroxyl means a -OH group. In this document, the term nitro means the -NO2 group.

The term lower, when used in conjunction with a chemical moiety such as acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy, means groups where ten or less, preferably six or less, non-hydrogen atoms are present in the substituent. Lower alkyl, for example, means an alkyl group which contains ten or less, preferably six or less carbon atoms. In some embodiments, the acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents as defined herein are, respectively, lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether present or not. alone or in combination with other substituents, such as the mentioned hydroxyalkyl and aralkyl (in such a case, for example, the atoms in the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

The term substituted means moieties having substituents replacing hydrogen atoms on one or more carbon atoms in the main chain. It should be understood that the terms substitution or substituted include the implied condition that such substitution is in accordance with the allowable valency of the substituted atom and substituent, and that the substitution results in a stable compound, for example, which does not undergo spontaneous transformation such as rearrangement, cyclization, elimination, and etc. As used herein, the term substituted includes all acceptable organic substituents. In a broad aspect, acceptable substituents include acyclic and cyclic, branched and straight, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. Allowable substituents may be one or more and be the same or different for the respective organic compounds. For the purposes of the present invention, heteroatoms such as nitrogen may have hydrogen substituents and/or any valid substituents of the organic compounds described herein that satisfy the valences of the heteroatoms. The substituents may include any of the substituents described herein, for example, halogen, hydroxyl, carbonyl (such as carboxyl, alkoxycarbonyl, formyl, or acyl), thiocarbonyl (such as a thioether, thioacetate, or thioformate), alkoxy, phosphoryl, phosphate, phosphonate, phosphinate, amino, amido, amidine, imine, cyano, nitro, azido, sulfhydryl, alkylthio, sulfate, sulfonate, sulfamoyl, sulfonamido, sulfonyl, heterocyclyl, aralkyl, or an aromatic or heteroaromatic moiety. Those skilled in the art will appreciate that the substituents may themselves be substituted if appropriate. Unless specifically indicated as being unsubstituted, chemical moieties referred to herein are to be understood as including substituted variants. For example, a referenced aryl group or moiety includes both substituted and unsubstituted variants.

The term thioalkyl as used herein means an alkyl group substituted with a thiol group.

The term thioester as used herein means a -C(O)SR ^x or SC(O)R ^x group, where R ^x is hydrocarbyl.

The term thioacid, thiocarboxylic acid or thiocarboxylic acid as used herein means a group represented by the formula -C(O)SH.

The term thiocarboxylate means a group represented by the formula -(C(O)S) ^- .

As used herein, a therapeutic agent that prevents a disorder or condition means a compound that, for a statistical subject, reduces the occurrence of a disorder or condition in a treated subject compared to an untreated control subject, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition. compared to an untreated control subject.

The term treatment includes prophylactic and/or therapeutic treatment. The term prophylactic or therapeutic treatment is known in the art and includes administering to the host one or more of the compositions in question. If the administration is prior to the clinical manifestation of the undesirable condition (e.g., a disease or other undesirable condition in the host animal), then the treatment is prophylactic (i.e., it protects the host from the development of the undesirable condition), while if the administration is after the manifestation of the undesirable condition, the treatment is therapeutic (i.e., it is intended to eliminate, mitigate or stabilize an undesirable condition or related side effects).

- 38 040967

The term prodrug encompasses compounds that are converted under physiological conditions to the therapeutically active agents of the present invention (eg a compound of formula (I)). A common method of obtaining a prodrug involves the inclusion of one or more selected fragments, which are hydrolyzed under physiological conditions, releasing the desired compound. In other embodiments, the prodrug is converted by enzymatic activity of the host animal. For example, esters or carbonates (eg esters or carbonates of alcohols or carboxylic acids) are the preferred prodrugs of the present invention. In some embodiments, some or all of the compounds of formula (I) in the composition above may be replaced by an appropriate suitable prodrug, where, for example, the hydroxyl in the parent compound is present as an ester or carbonate, or the carboxylic acid in the parent compound is present in ester form.

In this document, the term contain or containing in General is used in the sense of inclusion, in other words, allows the presence of one or more additional (unspecified) distinguishing features or components.

As used herein, the term including, as well as other forms such as include, includes, and included, are not intended to be limiting.

As used herein, the term amino acid means a molecule containing both an amino group and a carboxyl group, including the corresponding salts, esters, combinations of various salts, as well as tautomeric forms. In solution at neutral pH, the amino group and the acid group of an amino acid can exchange a proton, forming a doubly ionized, although generally neutral, fragment, defined as a zwitterion. In some embodiments, the amino acids are α-, β-, γ-, or δ-amino acids, including their stereoisomers and racemates. In this document, the term L-amino acid means an α-amino acid having a left-handed configuration with respect to the a-carbon, that is, a carboxylic acid of the general formula CH(COOH)(NH ₂ )-(side chain) having an L-configuration. Similarly, the term D-amino acid means a carboxylic acid of the general formula CH(COOH)(NH ₂ )-(side chain) having a dextrorotatory configuration relative to the α-carbon. The side chains of L-amino acids may include naturally occurring and non-naturally occurring fragments. Non-naturally occurring (ie, non-natural) amino acid side chains are fragments that are used in place of naturally occurring amino acid side chains, eg, in amino acid analogs.

The term amino acid residue as used herein means a fragment having a structure similar to the original amino acid.

An amino acid residue may be covalently bonded to another chemical moiety via the amino group of the residue or the carboxyl group of the residue (ie, the hydrogen atom -NH2 or -OH is replaced by a bond to another chemical moiety).

As used herein, the term amino acid side chain means a moiety that is covalently linked to a D- or L-amino acid structure and can be represented as CH(COOH)(NH2)-R. For example, in the case of alanine CH(COOH)(NH2)(CH ₃ ), the amino acid side chain (R) is —CH ₃ . Examples of an amino acid side chain include, but are not limited to, (C1-C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, or (C ₂ -C ₆ )alkynyl. The amino acid side chain may be substituted with one or more of the same or different substituents selected from, but not limited to, amino, amido, alkylamino, acylamino, carboxylic acid, carboxylate, thiocarboxylate, thioacid, hydroxy, cycloalkyl, (cycloalkyl)alkyl, aryl , heterocyclyl, heteroaryl, guanidino, -SH, -S(alkyl); where optionally cycloalkyl, aryl, heterocyclyl and heteroaryl are further substituted with one or more substituents such as hydroxy, alkoxy, halogen, amino, nitro, cyano or alkyl.

Amino acids include the twenty standard amino acids used in protein synthesis by most biological organisms. As non-natural amino acid residues, alpha and alpha-disubstituted amino acids, N-alkylamino acids, and naturally occurring amino acids substituted with lower alkyl, aralkyl, hydroxyl, aryl, aryloxy, haloalkyl, or acyl may be selected, but are not limited to.

For example, lysine can be substituted to form a non-natural amino acid, such as at the carbon atom of its side chain or, alternatively, by mono- or dialkylation of its NH ₂ end group (for example, when the side chain of the amino group of lysine, together with its substituents, forms a heterocyclic ring such as piperidine or pyrrolidine). In another example, the amino terminal side chain of lysine can form a ring with the amino acid backbone, as in capreomycidin. Additional non-natural lysine derivatives include homolysin and norlysin. The lysine side chain may alternatively be substituted with a second amino group. In the following example, the alkyl side chain portion of a lysine can be introduced into a carbocyclic ring structure to form a semi-rigid analog such as, for example, cyclohexyl or cyclopentyl.

- 39 040967

Throughout the description and claims of the present invention, the L-threonine residue and/or the L-threonine side chain mentioned in the compound of formula (I) and/or the corresponding preparations may be represented by any of the following formulas:

n n

L-threonine L-threonine L-threonine L-threonine

In some embodiments, the non-natural amino acid may be a derivative of a naturally occurring amino acid having one or more double bonds.

In other exemplary embodiments, in threonine, an ethyl, phenyl, or other higher alkyl group may replace the beta-methyl group. In histidine, the imidazole group may be substituted, or alternatively, the side chain alkylene skeleton may be substituted.

Additional examples of non-natural amino acids include homoserine and natural amino acid homologues.

In further exemplary embodiments, the non-natural amino acid may be alkylated (eg, methylated) at the alpha position.

Additional examples of non-natural amino acids include alpha, beta and beta, gamma dehydroamino analogs of amino acids.

Additional exemplary amino acids include penicillamine and betamethoxyvaline.

Additional examples of non-natural amino acids include amino acids in which the side chain includes amino, alkylamino, acylamino, -COO-alkyl, cycloalkyl, heterocyclyl, heteroaryl, guanidino, (cycloalkyl)alkyl, (heterocyclyl)alkyl, and (heteroaryl)alkyl.

A modified N-terminal amino group and a modified C-terminal carboxyl group mean that said amino group or carboxyl group is modified.

The modification of the N-terminal amino group preferably has the general formula -NRxRy, in which Rx is hydrogen or alkyl and Ry is alkyl, alkenyl, -C(=NH)NH2, alkynyl, acyl, cycloalkyl, aryl, or heterocyclyl.

Examples of N-terminal modifications include, but are not limited to, acetylated, formylated, or guanylated N-terminal groups.

The modification of the C-terminal carboxyl group preferably has the general formula COR _z (R _z replaces the hydroxyl group of the last amino acid); where R _z is NR ₇ R ₈ , alkoxy, amino or imide. The C-terminal carboxyl group can also be converted to a heterocyclic ring (such as a 1,2,4-oxadiaxol or 1,3,4-oxadiazole ring) optionally substituted with hydroxy, alkyl, hydroxyalkyl, alkoxyalkyl, or cycloalkyl.

The present invention includes pharmaceutically acceptable salts of the compounds of the invention and their use in the compositions and methods of the present invention. In some embodiments, contemplated salts of the present invention include, but are not limited to, alkyl-, dialkyl-, trialkyl-, or tetraalkylammonium salts. In some embodiments, provided salts of the present invention include, but are not limited to, salts that form L-arginine, benetamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc. In some embodiments, contemplated salts of the present invention include, but are not limited to, Na, Ca, K, Mg, Zn, or other metal salts.

Pharmaceutically acceptable acid addition salts may also exist as various solvates, for example with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such a solvate may be a crystallization solvent, a preparation or crystallization solvent, or a co-solvent.

Pharmaceutically acceptable means a material which is suitable for the preparation of a pharmaceutical composition, which is generally safe, non-toxic and not biologically or otherwise undesirable, and which is acceptable for veterinary pharmaceutical use as well as for human pharmaceutical use.

The term stereoisomers means any enantiomers, diastereoisomers or geometric isomers, for example, of the compounds of the present invention. When the compounds of the invention are chiral, they may exist in racemic or optically active form. Since the pharmaceutical activity of racemates or stereoisomers of compounds according to this

- 40 040967 The present invention may vary, it may be desirable to use compounds that are enriched in one of the enantiomers. In these cases, the final product or even intermediate compounds can be separated into enantiomeric compounds by chemical or physical means known to those skilled in the art, or even used as received in the synthesis. In the case of racemic amines, diastereomers are formed from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving reagents are optically active acids such as the R and S forms which have tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g. N-benzoylproline or N -benzenesulfonylproline) or various optically active camphorsulfonic acids. In addition, the chromatographic separation of enantiomers using an optically active resolving agent (such as, for example, dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chirally derivatized methacrylate polymers immobilized on silica gel) is advantageous.

In some embodiments, the compounds of the present invention may be racemic. In some embodiments, the compounds of the present invention may be enriched in one enantiomer. For example, a compound of the invention may have more than 30% ee, 40% ee, 50% ee, 60% ee, 70% ee, 80% ee, 90% ee, or even 95% ee or more. In some embodiments, the compounds of the present invention may have more than one stereocenter. In certain such embodiments, the compounds of the invention may be enriched in one or more diastereomers. For example, a compound of the present invention may have more than 30% de, 40% de, 50% de, 60% de, 70% de, 80% de, 90% de, or even 95% de or more.

The term subject includes mammals (particularly humans) and other animals such as domestic animals (eg, companion animals, including cats and dogs) and non-domestic animals (such as wild animals).

Naturally occurring amino acids are defined throughout the description and claims by traditional three-letter abbreviations, which are presented in table. 1 below.

Table 1

Amino acid codes

Name Three-letter Name Three-letter Alanine Ala Lysine Lys Arginine Arg Methionine Met Asparagine Asn Phenylalanine Phe Asparaginova asp Proline Pro Glutamine Glu Serene Ser Glutamine gin Threonine Thr G istidine His Tyrosine Tyr Isoleucine He Valine Vai

Abbreviations used throughout the description are summarized below by means of their specific meanings.

°С - degree Celsius;

% - percent;

saline - NaCl solution;

CH ₂ C1 ₂ /DCM - dichloromethane;

Boc - tert-butyloxycarbonyl;

Bzl - benzyloxycarbonyl;

Cs ₂ CO ₃ - cesium carbonate;

DIC - N,N'-diisopropylcarbodiimide;

DIPEA - N,N-diisopropylethylamine;

DMF - dimethylformamide;

EtOH - ethanol;

Et ₂ NH - diethylamine;

Fmoc - 9-fluorenylmethyloxycarbonyl;

g - gram;

HOBt - 1-hydroxybenzotriazole;

h - hours;

HPLC - high performance liquid chromatography;

K ₂ CO ₃ - potassium carbonate;

- 41 040967

LC-MS - liquid chromatography-mass spectrometry;

liquid NH3 - liquid ammonia;

mmol - millimol;

M - molar;

µl - microliter;

ml - milliliter;

mg - milligram;

MS (ES) - electrospray mass spectrometry;

min - minute;

Na - sodium;

NaHCO ₃ - sodium bicarbonate;

NH2NH2-H2O - hydrazine hydrate;

NMM - N-methylmorpholine;

Na ₂ SO ₄ - sodium sulfate;

NH2OH-HCl - hydroxylamine hydrochloride;

PD-1/PD-1 - programmed cell death 1;

PD-L1 - programmed cell death ligand 1;

PD-L2 - programmed cell death ligand 2 1;

prep-HPLC/preparative HPLC - preparative high performance liquid chromatography;

TEA/Et ₃ N - triethylamine;

TFAA - trifluoroacetic anhydride;

TLC - thin layer chromatography;

THF - tetrahydrofuran;

TIPS - triisopropylsilane;

TFA - trifluoroacetic acid;

t _R - retention time;

Trt is trityl or triphenylmethyl; etc.

experimental part

The present invention provides methods for preparing compounds of formula (I) according to the procedures of the following examples using appropriate materials. Those skilled in the art will appreciate that known variations in the conditions and processes of the following preparation procedures may be used to prepare these compounds. In addition, using the procedures described in detail, a person skilled in the art will be able to obtain additional compounds according to the present invention.

The intermediates or starting materials required for the synthesis are commercially available (commercial sources such as Sigma-Aldrich, USA or Germany; Chem-Impex USA; G.L. Biochem, China and Spectrochem, India), or alternatively the indicated intermediates or starting materials can be obtained using known literary methods. The invention is described in more detail using specific examples.

Purification and characterization of compounds.

Analytical HPLC method.

Analytical HPLC was carried out on a ZIC HILIC 200 A° column (4.6x250 mm, 5 µm), flow rate 1.0 ml/min. Elution conditions used: buffer A - 5 mmol ammonium acetate, buffer B - acetonitrile, equilibrate the column with 90% buffer B and elute with a gradient of 90 to 40% buffer B over 30 minutes.

Preparative HPLC method.

Preparative HPLC was carried out on a SeQuant ZIC HILIC 200 A° column (10x250 mm, 5 µm), flow rate: 5.0 ml/min. Elution conditions used: Buffer A: 5 mmol ammonium acetate (adjusted to pH-4 with acetic acid), Buffer B: Acetonitrile, Equilibrate the column with 90% buffer B and elute with a gradient from 90% to 40% buffer B over 20 min.

LC-MS was performed on an AP1 2000 LC/MS/MS triple quadrupole mass spectrometer (Applied biosystems) with an Agilent 1100 series HPLC with a DAD G1315 B using a Mercury MS column or using an Agilent LC/MSD VL single quadrupole mass spectrometer with HPLC Agilent 1100 series with DAD G1315 B, using a Mercury MS column, or using a Shimadzu LCMS 2020 quadrupole mass spectrometer with a UFLC Prominence system with SPD-20 A DAD.

- 42 040967

Examples

Example 1. Synthesis of compound 1.

Stage 1a.

ECF/Et ₃ N , water ΝΗ3/ΤΓΦ

1a

Ethyl chloroformate (3.4 g, 31.3 mmol) and Et ₃ N (7.0 ml, 52.8 mmol) were added to a solution of compound 1a (5.0 g, 26.4 mmol) in THF (20 ml) and stirred at -20°C for 20 min. After 20 minutes, 25% aqueous ammonia solution (10 ml, 132.0 mmol) was added to the vigorously stirred anhydride and stirred at 0-5° C. for 30 minutes. The completeness of the reaction was confirmed by TLC analysis. The volatiles were evaporated under reduced pressure and partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ solution followed by citric acid and brine. The separated organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give 4.0 g of compound 1b.

LC-MS: 89.3 (M-Boc+H) ⁺ .

Stage 1b.

I TFAA.

NH2 I

BocHN y pyridine BocHN^CN

ABOUT

1b1С

Trifluoroacetic anhydride (8.4 g, 39.9 mmol) was added to a solution of compound 1b (5 g, 26.6 mmol) in pyridine (21.0 ml, 26.6 mmol) and stirred at room temperature for 2 h. The completeness of the reaction was confirmed by TLC analysis.

The volatiles were evaporated under reduced pressure and partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ solution followed by citric acid and brine. The separated organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give 3.5 g of compound 1c, which was used directly in the next step.

Stage 1c.

I nh ₂ oh.hci I

BocHN^CN K ₂ COz/EJUN ΒοοΗΝ γ ^{2 Ν} 'ΟΗ

1s 1d

Hydroxylamine hydrochloride (0.92 g, 13.2 mmol), water (0.5 ml) and potassium carbonate (1.8 g, 13.2 mmol) were added to a solution of compound 1c (1.5 g, 8.8 mmol ) in EtOH (20 ml) and stirred at 86°C for 4 h. The reaction was confirmed complete by TLC analysis. The volatiles were evaporated under reduced pressure and partitioned between water and ethyl acetate. The organic layer was washed with brine, dried over Na ₂ SO ₄ , then filtered and evaporated under reduced pressure to give 0.9 g of compound 1d.

LC-MS: 104.3 (M-Boc+H) ⁺ .

Stage 1d.

I MCHaOCH^NSFycH^ V-\ _lul:

FmocHN --------------------* N-n NHFmoc

II 2. Compound 1d, NaOAc, ^υ ° AcOH 1e

Deoxo-Fluor (3.7 g, 16.1 mmol) was added to a solution of Fmoc-Ala-OH (5.0 g, 15.0 mmol) in CH ₂ Cl ₂ (50 ml) and stirred at 0°C for 1 h Then CH ₂ Cl ₂ was evaporated and triturated with hexane. Sodium acetate (0.2 g, 2.4 mmol) and compound 1d (0.5 g, 2.4 mmol) in acetic acid were taken in a round flask and stirred for 30 min. To this mixture was added Fmoc-Ala-COF (0.996 g, 3.1 mmol) and stirred at room temperature for 30 minutes. The reaction mixture was stirred at 90° C. for 3 hours. The completeness of the reaction was confirmed by TLC analysis. The volatiles were evaporated under reduced pressure and partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ solution followed by citric acid and brine. The separated organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give a residue. The residue was purified by silica gel column chromatography (eluent: 0-5% ethyl acetate in hexane) to give 0.3 g of compound 1e.

LC-MS: 379.0 (M-Boc+H) ⁺ , 501.3 (M+Na) ⁺ .

- 43 040967

Stage 1e.

Diethylamine (1.0 ml) was added to a solution of compound 1e (0.35 g, 0.73 mmol) in CH2C12 (10 ml). The mixture was stirred at room temperature for 3 hours, and then the resulting solution was concentrated under nitrogen and washed with diethyl ether and centrifuged to give 0.18 g of compound 1f, which was directly used in the next step.

Stage 1f.

The formation of the urea group was carried out using a combination of compound 1f (0.18 g,

0.71 mmol) in DMF (5.0 ml) with compound 1h (0.24 g, 0.77 mmol) at room temperature. The combination was initiated by adding TEA (0.2 g, 1.4 mmol) and the resulting mixture was stirred at room temperature. After completion of 3 hours the reaction mass was divided between water and ethyl acetate. The organic layer was washed with water followed by brine and dried over Na ₂ SO ₄ . The separated organic layer was filtered and evaporated under reduced pressure to give 0.2 g of compound 1g.

LC-MS: 450.1 (M+Na)+.

Stage 1g.

To a solution of compound 1g (0.15 g, 0.35 mmol) in CH ₂ Cl ₂ (1 ml) was added trifluoroacetic acid (0.5 ml) and a catalytic amount of triisopropylsilane and stirred at room temperature for 3 h. The resulting solution was concentrated under reduced pressure to obtain 0.10 g of a crude compound. The crude solid was purified as described under experimental conditions for the HPLC method.

LC-MS: 272.2 (M+H) ⁺ ; HPLC (t _R ): 6.2 min.

Synthesis of compound 1h.

Pyridine (12.0 g, 25.2 mmol) was added to a solution of H-Ala-OtBu (2.3 g, 12.6 mmol) in CH ₂ Cl ₂ (20 ml) and the resulting solution was stirred at room temperature for 5 -10 min. A solution of 4-nitrophenyl chloroformate (2.8 g, 13.8 mmol) in CH ₂ Cl ₂ (20 ml) was added to the above mixture and stirring was continued at room temperature for 1 h. The reaction was confirmed complete by TLC analysis. After completion of the reaction, it was diluted with CH ₂ Cl ₂ (50 ml) and washed with 1.0 M sodium bisulfate solution (50 mlx2) followed by washing with 1.0 M sodium carbonate solution (50 mlx2). The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give a crude compound, which was purified by silica gel column chromatography (eluent: 0-20% ethyl acetate in hexane) to give 2.0 g of compound 1h.

The compounds below were prepared in a manner analogous to that described in Example 1 (compound 1) with appropriate changes in reagents or amino acids, solvents, reagent amounts and reaction conditions. The analytical data of the compounds are shown in the table below.

- 44 040967

connection no. Structure zhhms (M+N) ⁺ HPLC (tR min) 2. °γ ^0Η νη ₂ P oA _ο γ° ^Η ^YYn ^A _N A ^H gL—on ^{n o} 403.2 12.6 3. NH ₂ I ⁿ % A ₀ A M ^{H H} S 373.1 9.4 4. νη ₂ °γ ^{0Η n} % A o A L—i ^{n n} έ 389.1 13.1 5. ΝΗ ₂ |^ΝΗ ₂ ⁿ % A o g ΑνΑν N—O ^{H H} O 388.1 21.9 6. BUT. \ /OH A g \ o u η/ _Ύ Υ?7 _Ύ ^οη ill— ^on o 344.2 12.4

Example 2. Synthesis of compound 7.

Stage 2a.

Compound 2a was synthesized using a similar procedure as shown in steps 1a-1c of example 1 (compound 1) using Boc-Tyr(tBu)-OH instead of Boc-Ala-OH to give 9 g of compound 2a.

Stage 2b.

HOBt (3.45 g, 30.8 mmol) and DIC (4.5 ml, 30.8 mmol) were added to a solution of Fmoc-Pro-OH (8.6 g, 25.6 mmol) in DMF (250 ml) at 0°C and stirred for 30 min. Compound 2a (9 g, 25.6 mmol) was further added to the above reaction mixture at the same temperature, and stirring was continued for 2 h at 0° C. followed by washing for 2 h at room temperature. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was quenched with ice water, the precipitated white solid was filtered, washed with water (1 L) and dried under reduced pressure. The solid was stirred with diethyl ether (500 ml) for 15 min, filtered

- 45 040967 and dried to obtain 14 g of compound 2b. LC-MS: 671.3 (M+H)+, 693.3 (M+Na)+. Stage 2c.

To a solution of compound 2b (13 g, 19.4 mmol) in acetonitrile (130 ml) was added acetic acid (10.0 ml) at room temperature and refluxed at 85°C for 12 h. using TLC analysis. The volatiles were evaporated under reduced pressure to give a crude semi-solid which was diluted with water and ethyl acetate. The organic layer was washed with NaHCO ₃ solution followed by citric acid and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give a crude solid which was diluted with 10% acetonitrile in hexane (500 ml) and stirred for 2 h to give a white solid. The white solid was filtered and washed with n-pentane (500 L) and dried to give 13 g of compound 2c.

LC-MS: 653.4 (M+H)+, 675.6 (M+Na)+.

Stage 2d.

‘BuO^^ ‘BuO.

χχXX

N '20% ^pipe R ^& D ^inv DXMN

BocHN^^ ^^NFmoc BocHN^^Yj

N--Ο NO

2c2d

Compound 2c (13g, 19.9 mmol) was added to a solution of 20% piperidine in DCM (150 ml) at 0°C and stirred at 0°C for 1 h. The reaction was confirmed complete by TLC analysis. The reaction mixture was concentrated under reduced pressure and diluted with hexane, stirred and filtered. The filtered solid was dissolved in EtOAc and washed with sat. NaHCO ₃ , brine, dried over Na ₂ SO ₄ , filtered and evaporated to give white compound 2d.

LC-MS 431.1 (M+H)+; 453.4 (M+Na)+.

Stage 2e.

DIPEA was added to a solution of compound 2e (5 g, 11.6 mmol) and compound 2d (5.3 g, 11.6 mmol) in dry THF (50 ml) at 0°C and stirred for 2 h. stir at room temperature for an additional 4 hours. The volatiles were evaporated and partitioned between ethyl acetate and water. The organic layer was washed with saturated NaHCO ₃ 10% citric acid, brine, dried over Na ₂ SO ₄ and concentrated under reduced pressure. The crude compound was purified by neutral alumina column chromatography using 25% ethyl acetate in hexane to give compound 2f.

LC-MS: 772.5 (M+Na)+.

Stage 2f.

A solution of compound 2f (6.5 g, 8.7 mmol) and trifluoroacetic acid (16 ml) in DCM (16 ml) was stirred at 0°C for 1 h. The resulting reaction mixture was evaporated under reduced pressure, diluted diethyl ether and filtered to obtain 4 g of the crude compound. The crude solid was purified using the preparative HPLC method described under experimental conditions.

LC-MS: 482.3 (M+H)+; HPLC: tR= 12.92 min.

Synthesis of compound 2e.

This compound was synthesized using a similar procedure as shown for compound 1h in Example 1, using H-Tyr(tBu)-OtBu instead of H-Ala-OtBu.

The compounds below were prepared in a manner analogous to that described in Example 2 (compound 7) with appropriate changes in reagents or amino acids, solvents, reagent amounts and reaction conditions. The analytical data of the compounds are shown in the table below.

No. connections Structure HCMS (M+H) ⁺ HPLC (t _R , min) 8. nh ₂ Η ₂ Ν /? Voh H ₂ N\\ J ^N ~° H y ^oH o 402.3 11.87 9. 1 ΐ ^Η2 \ oA ο γ ^ΟΗ /А /Ν. /A /OH Ν---Ο Ο 387.0 11.8 10. η ₂ ν^ο ο^οη /А /χ yun YY Ν---Ο Ο 428.9 9.2 AND. ι^ΝΗ ₂ ^but h < 0 γ ^0Η hAWSV Ν--0 ^{π π} 0 375.1 10.6 12. Η ₂ Ν^^ι |^ΝΗ ₂ ^^^νη ₂ hX\aaV ^h Ν--Ο ^{Π Π} Ο 443.2 - 13. νη ₂ ΑΑ _Ο Α ο γ ^ΟΗ hAvAV Ν—Ο ^{Η Η} Ο 421 11.0 14. But. _ο Ύ>Η η^ΑύΑΑΑυ II 1 η η and Ν--Ο Ο 394.3 7.6 15. νη ₂ . οΑ _ο γ ^ΟΗ Λγ/Af Ν--Ο ^Π Ο 345.3 12.7

- 47 040967

16. n°. A _o UON nLuOA™ L—A ^{n n} A 360.3 8.4 17. ^H O \u003d o L-A ^{n n} A 318.0 from 18. o ⁿ % η ₂ ν^- o OuALHO ¹ J ¹ —A ^{n n} A 331.0 12.5 19. XX. ^Η °ν\ o (XX AW/* Ν--O o 498.4 7.6 20. HATKAH N--O O 496.3 5.6 21. XX. AoV ^OH h ₂ nVX%V ^H Ν--O O 420.3 7.7 22. Owon ow° ⁿ N--O ^{M M} O 450.4 I,4 23. OH OH o^\ A o ¢^0 ΑγΑΛο L-A ⁿ A 386.1 14.5

- 48 040967

24. °γ ^0Η OH n X η ₂ ν γ γ Ν Ν γ Ν--0 0 400.0 14.0 25. Χα Ο °γΌΗ n/uuLA, ν—o AU 325.7 10.3 26. L o d ^he Ν--O ^{and and} Ο 499.3 9.4 27. ^Η % L ⁿ ο γ ^ΟΗ n ₂ Ay _g AA _g ^0H Il 1 η η I Ν--0 0 417.3 13.1 28. °^/ ^ΝΗ 2 νη ₂ no. D \ J A ° \u003d ⁰ f xD. /N. Αχ χΧ χΧ χΟΗ Η ₂ Ν AG Ar Ν Ν ΑΑ II 1 η η II Ν---0 0 387.9 14.9 29. 0^ /ΝΗ ₂ νη ₂ Α^ L \ jun ° 1 = ⁰ xxL ,Ν. xL Dx Dx χ ^ΟΗ Η ₂ Ν V ν ν Α< II 1 η η II Ν---Ο Ο 401.3 14.0 thirty. ΗζΝ^ΝΗ χΝΗ Γ ϊ 1 ⁰ ^ ^0Η Dx.x'^xDx .Ax -Ax.x- ^on η ₂ ν y nn AA Ν---0 0 416.2 13.4

- 49 040967

31. | ^ ^ NH ₂ NH ₂ L / OH SCH. AA OH N --- O O 388.1 AND 2 32. I Mz IZ iz / O=\ o o I 431.2 18.9 33. H ₂ N ^ NH NH O ^ NH ₂ / J o Y ^OH / \ JL / A / OH * Ai yA N --- 0 0 444.4 12.2 34. <TL s ill ^H ° ^A - ° Ύ ^ΟΗ η ₂ νΑΥν ^Λ νΑ ^οη J—i n n l 412.2 15.4 35. <1 1 ^Η2Ν Ί nsgA o 4 D- /L/OH ^η ΆΑβ yA Ν---Ο Ο 439.3 I,5 36. /\ Ο^/ΝΗ ₂ b ζ ^0Η / Υ Α\ Jk /A. .OH ^ΗίΝ Ϊ ^h Gy y G Ν---0 0 402.4 I,9 37. /\ ο<. .ON < ^nh ₂ Vb ^ ^on A/CH Jk A/ ^on _Η2Ν γ γ Ai A N---0 0 403.1 8.5

- 50 040967

38. τ О τ \ Ο 5=0 / ΖΙ Η °£ ζ I ζ '—' τ. MS Τ 417.0 8.4 39. ΝΗ ₂ ΑΖ ⁿ % A ο nAagAAa A A ⁿ A 407.5 12.2 40. nh ₂ \ \ /OH A o y^ χΑ/Ά/Α A A/ ^{he H} \u003d ^N A Ai d A N --- O O 373.1 12.7 41. ZA /u NH ₂ ^H \A L ^on JAx LAon ΗΚγ ^A N NX II 1 n n II Ν---0 0 460.2 11.9 42. But. ABOUT-. A o y ° ^{n Η} \u003d ^Ν A Ai yA Ν---0 O 437.2 12.9 43. 0 ^but \ ^ΗΝ \Α\_ _o ύ°η η ₂ νΑ ^Ν Αν ^Λ νΑ ^0Η II 1 n n II Ν---0 Ο 433.3 11.6 44. ON \ / \ .ON Αν A A Aon u^AAi yA Ν---Ο 0 346.5 11.9

- 51 040967

45. X about /ZX £°\z '----' Z οι X 400.1 10.6 46. nh ₂ he o^S. o.n. Αχ /L D. .OH ^H2N t Ty ychG N---0 0 419.0 12.7 47. H.O. HO\ θ .OH /L /NA /I OH ^H * ^N Y n^Y N---0 O 320.1 14.9 48. OH HO OH o^Ch o z^o D. /N. /A Jk / ^OH H,N Ti NN Tf II 1 η η II N---O O 404.1 14.7 49. Λ NH ₂ Α οΑ ογ“ H ₂ MAGUAAAA ^OH Ν--0 ^{Μ Μ} 0 385.1 12.6 50. ΑΑ νη ₂ ΤΑ ₀ Α _ο γΟΗ ^n'VAnAA''" Ν—0 ^{Η Η} 0 427.0 10.9 51. . ¹ I ζ Ο A τζ ΙΖ / Ο=< Ο \ Ζ ζζ δ 493.3 13.8 52. ψ 0 γ ^{0Η HzN} a TA i g N---O 0 386.1 8.5

- 52 040967

53. χϋΟ ^Η % οΑ ον ^ΟΗ Μ ^{Η Η} S 495.0 AND 2 54. νη ₂ 4 οΑ 0 γ ^0Η ,ΑΑλ'Υ Ν—Ο ^{Η Η} 0 359.0 14.0 56. ΝΗ ₂ ΗΝ' νη ₂ ο Aw οΑ^ q |Α ₀ Α /Ά A /A /He Η ₂ Ν Αγ ν ν Α< II 1 η η II Ν---0 0 401.0 13.6 58. νη ₂ HS. \χ ^ΟΗ ΑαΑΑ Ν---Ο Ο 374.9 14.1 59. ρ _Ν Η ₂ °γ ^ΝΗ2 ι = ° Ύ° ^Η D. / a /A JU xL /He ^Η > ^Ν Ar *mA yAr Ν---O O 416.1 12.9 60. ^N °\^\ nouA _oA Ν---O ⁿ 4 420.1 10.6 62. I o γ° X <1 o γζ / Ζ cm X 403.9 9.2

- 53 040967

63. ι^ΝΗ ₂ OH hXvaXt N--O ^{Μ Μ} O 403.1 14.8 64. Η/χ_Ν. ^ΝΗ he < νη ₂ Ж L νη ₂ X ο γ ² /4 JL D. .on η ₂ ν >< II 1 η η II Ν---Ο Ο 458.5 11.8 65. >οη ο ο 814.1 14.9 73. νη ₂ ο^- Ο f Η ₂ Ν^ν-Ο ^{Η Η} ο 391.6 - 74. Η ₂ Ν Ν-θ Ο 386.1 -

Example 3 Synthesis of Compound 75

n

BUT

he h ₂ n N----O O compound 75

The specified compound was synthesized using a similar procedure as shown in example 2, using compound 3b (obtained in accordance with the procedure below) instead of Fmoc-Pro-OH. The crude solid was purified using preparative HPLC as described under experimental conditions.

LC-MS: 417.5 (M+H)+, HPLC: t _R = 12.2 min.

Synthesis of compound 3b.

Stage 3 a.

_n O H ₂ N

Fmoc' ^N vAd'Bu HATU/ DIPEA/DMF

ΟΌΗ

0°С rt, 1.5 h

H ⁰

Fmoc' ^N X^O'Bu

Behind

Propylamine (2.9 ml, 35.25 mmol), HATU (14.8 g, 38.8 mmol) and DIPEA (12.3 ml, 70.5 mmol) were added to a solution of Fmoc-Glu-OtBu (15.0 g, 35.3 mmol) in DMF (50 ml) at 0° C. and stirred at room temperature for 1.5 h. Completion of the reaction was confirmed by TLC analysis. The reaction mixture was quenched with water, the resulting solid was filtered, washed with diethyl ether, dried under reduced pressure to give compound 3a (19.0 g) and used in the next step without purification.

LC-MS: 489.4 (M+Na+H) ⁺ .

- 54 040967

Stage 3b.

To a solution of compound 3a (19.0 g, 40.72 mmol) in trifluoroacetic acid (19.0 ml) was added a catalytic amount of triisopropylsilane and stirred at room temperature for 4 h. The resulting solution was evaporated under nitrogen, washed with diethyl ether to obtain 13.3 g of the crude compound, which was used in the next step without purification.

LC-MS: 417.5 (M+H)+.

The compounds below were prepared in a manner analogous to that described in Example 3 with appropriate changes in reagents or amino acids, solvents, reagent amounts and reaction conditions. The analytical data of the compounds are shown in the table below.

connection no. Structure LCMS (M+N) ⁺ HPLC (t _R , min) 76. ^H % to ₀ uOH ΑνΛν N--O ^M O 459.3 9.8 77. ^Η °Ί _p /Ν = π T ^Η * ^Ν A An AA<on Ν-Ο Η H 445.0 9.6 78. Ο^ΝΗ ₂ BUT. [ J A and A % ^he 587.3 10.7

Example 4 Synthesis of Compound 79

connection 179

The specified compound was synthesized using a similar procedure as shown in example 2, using compound 4f (according to the procedure below) instead of H-Tyr(tBu)-OtBu. The crude solid was purified using preparative HPLC as described under experimental conditions.

LC-MS: 415.0 (M+H)+, HPLC: t _R = 13.1 min.

Synthesis of compound 4f.

Stage 4a.

Methyl iodide (4.9 g, 34 mmol) and potassium carbonate (6.0 g, 44 mmol) were added to a solution of compound 4a (7.1 g, 22 mmol) in DMF (70 ml) and stirred at room temperature for 3 h. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between ice water and ethyl acetate. The organic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give 6.4 g of compound 4b.

LC-MS: 324.0 (M+H) ⁺ .

- 55 040967

Stage 4b.

\.O'Bu \.O'Bu

T NoH _fi o T ^h

CbzHN γ MeOH CbzHN γ NH ₂ o O

4b 4c

Hydrazine hydrate (6.4 ml) was added to a solution of compound 4b (6.4 g) in methanol (64 ml) and stirred at room temperature for 12 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between ice water and ethyl acetate. The organic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give 5.5 g of compound 4c.

LC-MS: 324.2 (M+H)+.

Stage 4c.

\.O'wee \,O*wee

Y H HOBT/EDC.HCI T n V

I N -------------► YES ,Ο'Βυ

CbzHN γ 'NH ₂ DIPEA/DMF CbzHN γ N O o ⁿ

4s 4d

2-(tert-butoxy)acetic acid (1.16 g, 8.8 mmol), HOBT (1.3 g, 9.6 mmol), EDC.HCl (1.83 g, 9.6 mmol), DIPEA (2.5 g, 21 mmol) was added to a solution of compound 4c (2.6 g, 8.8 mmol) in DMF (50 ml) and stirred at room temperature for 4 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between ice water and ethyl acetate. The organic layer was washed with NaHCO ₃ , citric acid, brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give crude compound 6, which was further purified by silica gel column chromatography (eluent: 0-50% ethyl acetate in hexane) with obtaining 3 g of product 4d.

LC-MS: 438.2 (M+H) ⁺ .

Stage 4d.

Triphenylphosphine (6.5 g, 24 mmol), iodine (6.2 g, 24 mmol), TEA (3.5 g, 35 mmol) were added to a solution of compound 4d (3.1 g, 7.0 mmol) in THF (50 ml) and DMF (10 ml) and stirred at room temperature for 3 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between sodium thiosulfate solution and ethyl acetate. The organic layer was washed with brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give crude compound 4e, which was further purified by silica gel column chromatography (eluent: 0-50% ethyl acetate in hexane) to give 2.4 g of product 4e.

LC-MS: 420.2 (M+H) ⁺ .

Stage 4e.

\.Ο ^ι Βυ \-O ¹ Vi

Y Pd(OH) ₂ T

CbzHN'^Y \—MeOH H ₂ N'>^ \— nn oteu nV o'wee

4e 4f

Palladium hydroxide (0.5 g) was added to a solution of compound 4e (2.3 g) in methanol (30 ml) and stirred under H2 gas at room temperature for 2 h. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was filtered over a Celite pad and the filtrate was evaporated under reduced pressure to give 1.2 g of product 4f.

LC-MS: 286.1 (M+H) ⁺ .

Stage 4f.

4-Nitrophenyl chloroformate (0.85 g, 4.2 mmol), pyridine (0.36 g, 4.6 mmol) was added to the solution

- 56 040967 compounds 4f (1.2 g, 4.2 mmol) in CH ₂ CL ₂ (40 ml) and stirred at room temperature for 1 h. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between ice water and ethyl acetate. The organic layer was washed with citric acid brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give crude compound 4g, which was further purified by silica gel column chromatography (eluent: 0-40% ethyl acetate in hexane) to give 1.8 g product 4g.

Example 5 Synthesis of Compound 80

The specified connection was synthesized using a similar procedure as shown in example 2, using compound 5b (obtained in accordance with the procedure below) instead of H-Tyr(tBu)-OtBu. The crude solid was purified using preparative HPLC as described under experimental conditions.

LC-MS: 431.5 (M+H)+, HPLC: t _R = 12.6 min.

Synthesis of compound 5b.

Stage 5a.

To a solution of Fmoc-Thr(tBu)-OH (5.5 g, 13.8 mmol) in DMF (25.0 ml) K ₂ CO ₃ (2.9 g, 20.8 mmol) was added at 0°C and stirred for 30 minutes followed by the addition of iodopentane (2.1 ml, 16.6 mmol) at 0°C. The above reaction mixture was stirred at room temperature for 14 hours. Completion of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ , brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give compound 5a (6.86 g) and used in the next step without purification. LC-MS: 246.4 (M-Fmoc+H) ⁺ . Stage 5b.

20% piperidine in

CH ₂ C1 ₂ °C-rt, 1 h

ABOUT

5a 5b

To compound 5a (6.86 g, 14.67 mmol) was added 20% piperidine in CH ₂ Cl ₂ (34.3 ml) and the reaction mixture was stirred at room temperature for 1 h. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was evaporated under nitrogen and partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ , brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure. The crude compound 5b was purified by silica gel column chromatography (Eluent: 0-40% ethyl acetate in hexane LC-MS: 246.1 (M+H) ⁺ .

Example 6 Synthesis of compound 81.

to obtain 2.5 g of 5b.

The specified connection was synthesized using a similar procedure as shown in example 2, using compound 6b (obtained in accordance with the procedure below) instead of H-Tyr(tBu)-OtBu. The crude solid was purified using preparative HPLC as described under experimental conditions.

LC-MS: 443.8 (M+H) ⁺ , HPLC: t _R = 8.8 min.

- 57 040967

Synthesis of compound 6b.

Stage 6a.

To a solution of Fmoc-Gln(Trt)-OH (5.0 g, 8.2 mmol) in DMF (25.0 ml) HATU (3.4 g, 9.0 mmol), H-Thr(OtBu)-OtBu (1.9 g, 8.2 mmol) and DIPEA (2.9 ml, 16.4 mmol) were added at 0° C. and stirred at room temperature for 2 h. Completion of the reaction was confirmed by TLC analysis. The reaction mixture was quenched with water, the resulting solid was filtered, washed with hexane and dried to give 7.4 g of compound 6a.

LC-MS: 824.1 (M+H)+.

Stage 6b.

H

To compound 6a (7.4 g, 8.9 mmol) was added 20% piperidine in CH ₂ Cl ₂ (37.0 ml), the reaction mixture was stirred at room temperature for 2 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was evaporated under nitrogen and partitioned between water and DCM. The organic layer was washed with NaHCO ₃ , brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure. The crude compound was washed with hexane and dried under reduced pressure to obtain 3.7 g of compound 6b.

LC-MS: 601.8 (M+H) ⁺ .

Example 7 Synthesis of Compound 82

connection 82

The specified compound was synthesized using a similar procedure as shown in example 2, using compound 7c (obtained in accordance with the procedure below) instead of compound 2d. The crude solid was purified using preparative HPLC as described under experimental conditions.

LC-MS: 443.8 (M+H) ⁺ , HPLC: t _R = 8.8 min.

Synthesis of compound 7c.

Stage 7a.

EDC.HCl (2.9 g, 14.97 mmol), HOBt (2.3 g, 14.97 mmol), Fmoc-Gly-OH (2.97 g, 9.98 mmol) and NMM (2.7 ml, 24.95 mmol) was added to a solution of compound 7a (5.83 g, 9.98 mmol, The indicated compound 7a was synthesized using a similar procedure as shown for compound 2d in example 2) in DMF (30.0 ml) at 0° C. and stirred at room temperature for 18 hours. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with citric acid, NaHCO ₃ , brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure, further purified by silica gel column chromatography (eluent: 10% ethyl acetate in hexane-100% ethyl acetate) to give 6.0 d compound 7b.

LC-MS: 885.4 (M+Na+H) ⁺ .

- 58 040967

Stage 7b.

Diethylamine (24.0 ml) was added to a solution of compound 7b (6.0 g, 6.95 mmol) in CH ₂ Cl ₂ (24.0 ml). The reaction mixture was stirred for 6 h at room temperature and the resulting solution was concentrated under nitrogen to give compound 7c, which was purified by silica gel column chromatography (eluent: 0-10% ethyl acetate in hexane, then 2% methanol in DCM) to give 4.5 g of compound 7c.

LC-MS: 641.2 (M+H)+.

Example 8 Synthesis of Compound 83

hh ₂

WOD. ΝΗ/ΤΓΦ

About 8b

S2P5OOOS1/Ν Μ Μ o 8a

Stage 8a.

Ethyl chloroformate (2.35 ml, 25.05 mmol) and N-methylmorpholine (2.75 ml, 25.05 mmol) were added to a solution of compound 8a (5.0 g, 8.35 mmol) in THF (50 ml) and stirred at -20°C. In 20 minutes. an aqueous ammonia solution (3.0 ml) was added to the actively stirred anhydride formed in-situ and stirred at 0-5° C. for 1 hour. The reaction was confirmed to be complete by TLC analysis. The reaction mixture was poured into water and hexane, the resulting solid was filtered, washed with hexane and dried to obtain 4.2 g of compound 8b.

LC-MS: 596.3 (M+H) ⁺ .

Stage 8b.

Triethylamine (3.5 ml, 24.7 mmol) and trifluoroacetic anhydride (1.5 ml, 10.6 mmol) were added to a solution of compound 8b (4.2 g, 7.1 mmol) in THF (70 ml) and stirred at 0°C for 1 h. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ , brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure to give 4 g of crude compound 8c, which was used in the next step without further purification.

LC-MS: 578.2 (M+H) ⁺ , 600.2 (M+Na) ⁺ .

Stage 8c.

To compound 8c (1.5 g, 2.59 mmol) was added 20% piperidine in CH ₂ Cl ₂ (20.0 ml) and the reaction mixture was stirred at 0°C for 1 h. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was evaporated under nitrogen and partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ , brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure. The crude compound was washed with hexane and 20% diethyl ether in hexane to give

- 59 040967 with 0.8 g of compound 8d, which was used in the next step without further purification.

LC-MS: 378.4 (M+Na)+.

Stage 8d.

Triethylamine was added to a solution of compound 8d (0.8 g, 2.3 mmol) in DMF (15.0 ml) at 0° C. over 2 min. Compound 8i (1.0 g, 27.0 mmol) was then added. The reaction mixture was stirred at room temperature for 2 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was poured into water and the resulting solid was filtered, washed with hexane and 20% diethyl ether in hexane to obtain 1.2 g of compound 8e.

LC-MS: 613.6 (M+H) ⁺ .

Stage 8e.

nh ₂ oh.hci to ₂ cos/eyun/ water

Hydroxylamine hydrochloride (0.204 g, 29.37 mmol), water (10.0 ml) and potassium carbonate (0.4 g, 29.37 mmol) were added to a solution of compound 8e (1.2 g, 19.58 mmol) in EtOH (26.0 ml) and stirred at 85°C for 2 hours. The reaction was confirmed complete by TLC analysis. The reaction mixture was evaporated under reduced pressure and quenched with water, the resulting solid was filtered, washed with hexane and dried to obtain 1.0 g of compound 8f.

LC-MS: 646.9 (M+H) ⁺ .

Stage 8f.

DIC (0.29 ml, 18.58 mmol) and HOBt (0.25 g, 18.58 mmol) were added to a solution of Boc-Ser(tBu)-OH (0.4 g, 15.48 mmol) in DMF ( 20.0 ml) at 0°C and stirred for 30 minutes followed by the addition of compound 8f (1.0 g, 15.48 mmol) and stirred at room temperature for 1.5 hours. The reaction was confirmed complete by TLC analysis . The reaction mixture was quenched with water, the resulting solid was filtered, washed with diethyl ether, dried under reduced pressure to give 1.4 g of compound 8g, which was used in the next step without further purification.

LC-MS: 889.4 (M+H) ⁺ .

Stage 8g.

Acetic acid (1.2 ml) was added to a solution of compound 8g (1.2 g, 13.49 mmol) in 2-methyl THF (30.0 ml). The reaction mixture was stirred at 84° C. for 12 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed with NaHCO ₃ , brine, dried over Na ₂ SO ₄ and evaporated under reduced pressure. The crude compound was washed with hexane and 5% diethyl ether in hexane to give 0.45 g of compound 8h, which was used in the next step without further purification.

LC-MS: 871.4 (M+H) ⁺ .

- 60 040967

Stage 8h.

Compound 83

Triisopropylsilane (0.25 ml) and water (0.25 ml) were added to a solution of compound 8h (0.4 g, 4.55 mmol) in trifluoroacetic acid (9.5 ml) and stirred at room temperature for 2 h. The resulting solution was evaporated under nitrogen to give 0.1 g of crude compound 83. The crude solid was purified using the preparative HPLC method described under experimental conditions.

LC-MS: 361.1 (M+H)+; HPLC: t _R = 13.9 min.

Synthesis of compound 8i.

To a solution of HCl.H-Thr( ^t Bu)-O ^t Bu (8 g, 29.9 mmol) in CH ₂ Cl ₂ (80 ml) was added pyridine (5.9 g, 74.0 mmol) and the indicated solution stirred for 5-10 min at room temperature. To this was added a solution of 4-nitrophenyl chloroformate (7.2 g, 35.0 mmol) in CH ₂ Cl ₂ and the resulting mixture was stirred for 30 min at room temperature. The completion of the reaction was confirmed by TLC analysis. After completion of the reaction, the reaction mixture was diluted with CH ₂ Cl ₂ and washed with water and 5.0 M citric acid solution. The separated organic layer was dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to give a crude compound, which was purified by silica gel column chromatography (eluent: 0-20% ethyl acetate in hexane) to give 9 g of compound 8i.

LC-MS: 397.3 (M+H)+.

Example 9 Synthesis of Compound 84

Stage 9a.

To a stirred solution of compound 9a (1.00 g) in DCM (40.0 ml) was added pyridine (0.33 ml) followed by the addition of cyclopropanesulfonyl chloride (0.33 ml) and the resulting mixture was stirred at ambient conditions for 16 h, whereupon TLC analysis indicated completion of the reaction. The reaction mixture was partitioned between water and DCM and the organic layer was washed with dil. HCl (1.0 N). The organic phase was then dried over Na ₂ SO ₄ and concentrated under reduced pressure to give the crude sulfonamide, which was further purified by column chromatography (neutral alumina, eluent hexane-EtOAc (2:3)) to give the desired compound 9b (1.10 G).

LC-MS: 735.4 (M+H+H ₂ O)+.

- 61 040967

Stage 9b.

compound 94

The specified connection 84 was obtained from compound 9b in accordance with the procedure described in stage 1 g of example 1 with appropriate changes in reagents, amounts of reagents, solvents and reaction conditions.

LC-MS (M+H)+: 319.9

The compounds below were prepared in a manner analogous to that described in Example 9 (compound 84), with appropriate changes in reagents or amino acids, solvents, reagent amounts, and reaction conditions. The analytical data of the compounds are shown in the table below.

No. connections Structure HCMS (M+H) ⁺ 85. About H ₂ N^k 0 NO-. m L / \\ ' n ° η ₂ ν _ν ό 336.2 86. About H ₂ N^4 0 but—. _m 1 / \\ A H ⁰ H ₂ N N'O 308.3 87. O H ₂ N^_ 0 I BUT—. ... I \_/^r N % / \\ A H ° H ₂ N N'° 322.3 88. About H ₂ N^k 0 GU NO—. _m 1 \ x-O / v A n ⁰ Η ₂ Ν Ν' ⁰ 348.2 89. O Η ₂ Ν^- ο P no—. .. λ / \\ ' Η ° Η ₂ Ν ν-Ο 362.3 90. ^Η % H ₂ A ο ² JJ but 413.1

Example 10 Synthesis of Compound 91

compound 91

- 62 040967

Stage 10a.

DIPEA (0.11 g, 0.89 mmol) was added under an inert atmosphere to a stirred solution containing compound 10a (0.25 g, 0.4 mmol), carboxylic acid 10c (74 mg, 0.44 mmol) and HATU ( 0.23 g, 0.61 mmol) in dry DMF (3.0 ml). The reaction mixture was stirred at ambient conditions for 16 hours. The resulting reaction mixture was partitioned between water (50 ml) and EtOAc (25 ml) and the aqueous layer was extracted with EtOAc (25 mlx3). The combined organic phases were washed once with cold water, dried over Na ₂ SO ₄ and the solvent was removed under reduced pressure to give the title compound 10b (0.12 g).

LC-MS: 779.0 (M+H+H ₂ O)+.

Stage 10b.

ABOUT

The indicated compound 90 was obtained from compound 10b according to the procedure described in step 1g of example 1, with appropriate changes in reagents, amounts of reagents, solvents and reaction conditions.

LC-MS (M+H)+: 363.9

The compounds below were prepared in an analogous manner to that described in Example 10 (compound 91), with appropriate changes in reagents or amino acids, solvents, reagent amounts, and reaction conditions. The analytical data of the compounds are shown in the table below.

No. connections Structure zhhms (M+N) ⁺ 92. nh ₂ o ^ o HO ^ mA L H ₂ N N'O ^N HN- / 329.3 93. nh ₂ O^A o he ^but ~\ n^A AA / DH / |_| 2 H ₂ N n-0 NH ₂ 317.1

Example 11 Synthesis of Compound 94

- 63 040967

Stage 11a.

To a stirred solution of 11a (2.00 g) in DCM (40.0 ml) was added pyridine (0.53 ml) followed by 4-nitrophenyl chloroformate (0.65 g) and the resulting mixture was stirred at ambient conditions for 16 h The reaction mixture was further partitioned between water and DCM and the organic layer was washed with dil. HCl (1.0 N). The organic phase was then dried over Na ₂ SO ₄ and concentrated under reduced pressure to give the crude sulfonamide, which was further purified by column chromatography (neutral alumina, eluent with hexane EtOAc (2:3)) to give the desired carbamate 11b (2.66 g) .

LC-MS: 801.4 (M+H+Na)+.

Stage 11b.

Amine 11e (0.55 g, 4.09 mmol) was added to a stirred solution of compound 11b (2.66 g, 3.41 mmol) in ethanol (30.0 ml) and the resulting mixture was refluxed at 75°C in for 16 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was evaporated under reduced pressure, diluted with ethyl acetate (75 ml) and washed successively with water (80 ml) followed by washing with us. K2CO3 solution (50 ml) and brine (50 ml). After drying organic substances Na ₂ SO ₄ ; the solvents were removed under reduced pressure to give the title compound 11c (1.0 g).

LC-MS: 775.3 (M+H) ⁺ .

Stage 11c.

ABOUT

The indicated compound 91 was obtained from compound 11c according to the procedure described in step 1g of example 1, with appropriate changes in reagents, amounts of reagents, solvents and reaction conditions.

LC-MS (M+H)+: 377.2.

Example 12 Synthesis of Compound 95

- 64 040967

Stage 12a.

Acetic acid (0.2 ml) was added to a stirred solution of compound 12a (500 mg) and compound 12d (134 mg) in dry DCE (20.0 ml) and the resulting mixture was stirred at room temperature for 12 h. Sodium borohydride (77 0 mg) was added to the above mixture and the resulting mixture was stirred at room temperature for 4 hours. The reaction mixture was diluted with water (25 ml) and extracted with DCM (3x25 ml). The combined organic phases were washed with brine and water, dried over Na ₂ SO ₄ and the solvents were removed under reduced pressure to give crude compound 12b, which was taken to the next step without further purification (200 mg).

LC-MS: 708.0 (M+H)+.

Step 12b.

The title compound 92 was prepared from 12b according to the procedure described in step 1g of Example 1 with appropriate changes in reagents, reagent amounts, solvents and reaction conditions.

LC-MS (M+H) ⁺ : 310.2

Example 13 Synthesis of Compound 96

Stage 13a.

Compound 13d (77.0 mg) was added to a stirred solution of compound 13a (0.3 g) in toluene (10.0 ml) under an inert atmosphere, and the resulting mixture was heated at 60° C. for 16 h. The solvent was removed under reduced pressure, the crude product was repeatedly washed with pentane (2x10 ml) and diethyl ether (2x10 ml) and dried under reduced pressure to obtain compound 13b (200.0 mg).

LC-MS: 702.0 (M+H-'Bp) ⁺ .

Stage 2

The title compound 96 was prepared from 13b according to the procedure described in step 1g of Example 1 with appropriate changes in reagents, reagent amounts, solvents and reaction conditions.

LC-MS (M+H) ⁺ : 360.0.

- 65 040967

Example 14 Synthesis of Compound 97

compound 97

Stage 14a.

O s' ^CI OH SO ₂ CI ₂ '°

O pyridine, Et ₂ O OrM^^ ² -78°C to RT, 4 h 14a

A solution of 4-nitrophenol (1.3 g, 9.99 mmol) and pyridine (0.8 ml, 9.99 mmol) in Et2O (20 ml) was added dropwise to a solution of SO ₂ Cl ₂ (0.8 ml, 9 .99 mmol) in Et ₂ O (20 ml) at -78°C in an argon atmosphere. The reaction mixture was allowed to warm to room temperature, stirred for 4 hours. The completeness of the reaction was confirmed by TLC analysis. The volatiles were evaporated under reduced pressure to give a crude compound. The crude compound was purified by silica gel column chromatography (eluent: 0-3% ethyl acetate in hexane) to give 1.2 g of compound 14a.

Ή NMR (400 MHz, CDCl ₃ ): δ 8.39-8.36 (m, 2H), 7.61-7.57 (m, 2H).

Step 14b.

A mixture of compound 14b (0.6 g, 2.59 mmol), molecular sieves (1.0 g), 4-nitrophenol (0.72 g, 5.18 mmol) and Et ₃ N (1.1 ml, 7 77 mmol) in dry CH ₂ Cl ₂ (25.0 ml) was added dropwise to a solution of compound 14a (1.2 g, 5.18 mmol) in dry CH ₂ Cl ₂ (5.0 ml) at -78° C in an argon atmosphere. After stirring for 30 minutes, the reaction mixture was allowed to warm to room temperature over 2 hours. The completeness of the reaction was confirmed by TLC analysis. The reaction mixture was evaporated under reduced pressure to give a crude compound. The crude compound was purified by silica gel column chromatography (eluent: 0-7% ethyl acetate in hexane) to give 0.7 g of compound 14c.

1H NMR (300 MHz, CDCl ₃ ): δ 8.30-8.27 (m, 2H), 7.52-7.49(m, 2H), 5.70-5.67 (1H d, J 9 .6), 4.17-3.90(1H, m), 1.49(9H, s), 1.28-1.23(3H, m), 1.15(9H, s).

Stage 14c.

Compound 14c (0.69 g, 1.59 mmol) in THF (5.0 ml) was added to a stirred solution of compound 14d (0.7 g, 1.14 mmol) and Et ₃ N (0.5 ml, 3, 42 mmol) in dry THF (10.0 ml) and the resulting reaction mixture was stirred at 70°C for 3 hours. The completeness of the reaction was confirmed by TLC analysis. The volatiles were evaporated under reduced pressure to give a crude compound. The crude compound was purified by silica gel column chromatography (eluent: 0-33% ethyl acetate in hexane) to give 0.55 g of compound 14e.

LC-MS: 907.4 (M+H)+.

- 66 040967

Stage 14d.

compound 97

Triisopropylsilane (0.25 ml) and water (0.25 ml) were added to a solution of compound 14e (0.55 g, 0.55 mmol) in trifluoroacetic acid (9.5 ml) and stirred at room temperature for 2 h. The resulting solution was evaporated under nitrogen to give 0.3 g of crude compound 97. The crude solid was purified using the preparative HPLC method described under experimental conditions.

LC-MS: 397.0 (M+H)+; HPLC: t _R = 10.547 min.

The compounds below were prepared in a manner analogous to that described in Example 14 (compound 97), with appropriate changes in reagents or amino acids, solvents, reagent amounts, and reaction conditions. The analytical data of the compounds are shown in the table below.

Although the present application has been illustrated by some of the previous examples, it is not limited to them; on the contrary, the present application covers the general area as described above. Various modifications and implementations may be made without departing from their spirit or scope. For example, the following compounds, which can be obtained by following a similar procedure described above, with suitable modification known to a person skilled in the art, are also included in the scope of this application.

- 67 040967

- 68 040967 nant PD-L1.

Example 15 Retention of the Proliferating Ability of Mouse Splenocytes in the Presence of Recombi Recombinant mouse PD-L1 (rm-PDL-1, catalog number: 1019-B7-100; R&D Systems) was used as a source of PD-L1.

Requirements.

Mouse splenocytes obtained from C57 BL6 mice aged 6 to 8 weeks; RPMI 1640 (GIBCO, catalog number: 11875); high glucose DMEM (GIBCO, catalog number: D6429); fetal bovine serum [Hyclone, catalog number: SH30071.03]; penicillin (10,000 units/ml) - streptomycin (10,000 µg/ml) in solution (GIBCO, catalog number: 15140-122); sodium pyruvate solution in modified Eagle's medium (MEM) 100 mM (100x) (GIBCO, catalog number: 11360); non-essential amino acid (GIBCO, catalog number: 11140); L-glutamine (GIBCO, catalog number: 25030); anti-CD3 antibody (eBiosciences - 16-0032); anti-CD28 antibody (eBiosciences 16-0281); ACK lysis buffer (1 ml) (GIBCO, catalog number: A10492); Histopaque solution (density 1.083 g/mL) (SIGMA 10831); trypan blue solution (SIGMA-T8154); 2 ml Norm Ject luer lock syringe (Sigma 2014-12); nylon cell sieve 40 µm (BD FALCON 35230); hemacytometer (Bright line-SIGMA Z359629); FACS buffer solution (PBS/0.1% BSA): phosphate buffered saline (PBS) pH 7.2 (Hi-Media TS1006) with 0.1% bovine serum albumin (BSA) solution (SIGMA A7050) and sodium azide ( SIGMA 08591); 5 mM CFSE stock solution: CFSE stock solution was prepared by diluting lyophilized CFSE with 180 µl of dimethyl sulfoxide (DMSO C2H6SO, SIGMA-D-5879) and placing aliquots in tubes for later use. Working concentrations were titrated from 10 μM to 1 μM. 0.05% trypsy solution

- 69 040967 on (eBioscience-650850-85) and 0.02% EDTA solution (SIGMA 59417C); 96-well ELISA plates (Corning CLS3390); Caliber BD FACS (E6016); recombinant murine B7-H1/PDL1 Fc Chimera, (rm-PD-L1 catalog number: 1019-B7-100).

Protocol.

Obtaining and cultivation of splenocytes.

Splenocytes were harvested in a 50 ml Falcon tube by mashing mouse spleen on a 40 μM cell sieve and then treated with 1 ml of ACK lysis buffer for 5 minutes at room temperature. After washing with 9 ml of RPMI complete growth medium, the cells were resuspended in 3 ml of 1xPBS in a 15 ml tube. 3 ml of Histopaque was carefully added to the bottom of the tube, leaving the top layer of the splenocyte suspension intact. After centrifugation at 800xg for 20 minutes at room temperature, the opaque layer of splenocytes was carefully collected without disturbing/mixing the layers. Splenocytes were washed twice with cold 1xPBS, and then the total number of cells was counted using exclusion method using trypan blue, and used further in cell studies.

Splenocytes were cultured in complete RPMI medium (RPMI+10% fetal bovine serum+1 mM sodium pyruvate+10,000 units/ml penicillin and 10,000 (µg/ml streptomycin) and kept in a CO ₂ incubator containing 5% CO ₂ at 37°C.

Proliferation study using CFSE.

Carboxyfluorescein succinimidyl ether (CFSE) is a dye that passively diffuses into cells and binds to intracellular proteins. Harvested splenocytes (1x106 cells/ml) were treated with 5 μM CFSE in a prewarmed 1xPBS/0.1% BSA solution for 10 min at 37°C. Excess CFSE was quenched by adding 5 volumes of ice-cold culture medium to the cells and incubating on ice for 5 min. CFSE-labeled splenocytes were washed an additional three times with ice-cold complete RPMI medium. CFSE-labeled splenocytes (1x105) were plated in wells containing either MDA-MB231 cells (1x105 cells grown in high glucose DMEM) or recombinant human PDL-1 (100 ng/mL) and test compounds. Splenocytes were stimulated by adding anti-mouse CD3 and anti-mouse CD28 antibody (1 μg/ml each), and the culture was additionally incubated for 72 h at 37° C. in 5% CO ₂ medium. Cells were harvested and washed three times with ice-cold FACS buffer and % proliferation was analyzed by flow cytometry with excitation filters at 488 nm and emission at 521 nm.

Collection, processing and analysis of data.

The percentage of splenocyte proliferation was analyzed using the cellquest FACS program, and the percentage of preservation of splenocyte proliferative ability by the compound was evaluated by subtracting the % baseline proliferation value and normalizing to the % stimulated splenocyte proliferation (positive control), taken as 100%. The results are presented in table. 2.

Stimulated splenocytes: splenocytes+anti-CD3/CD28 stimulation Background proliferation: splenocytes+anti-CD3/CD28+PD-L1.

Compound Proliferation: splenocytes+anti-CD3/CD28+PD-L1+compound.

The effect of the compound was examined by adding the compound at the required concentration to anti-CD3/CD28 stimulated splenocytes in the presence of a ligand (PDL-1).

- 70 040967

table 2

Percentage of Splenocyte Proliferating Ability Retained for Compounds of the Invention

No. connections Splenocyte Proliferation Percentage Maintained (@ 100 nM) No. connections Splenocyte Proliferation Percentage Maintained (@ 100 nM) 1 76 39 75 2 68 40 67 3 73 41 78 4 44 42 50 5 38 43 60 6 69 44 71 7 91 45 17 8 48 46 53 9 84 47 49 10 84 51 20 eleven 66 52 71 13 62 56 26 14 92 60 99 15 28 62 35 16 50 75 119 17 58 76 67 18 57 77 75 19 54 78 42 20 51 79 37 21 51 80 55 23 49 81 18 24 17 82 12 27 37 83 34 28 36 84 71 thirty 65 88 10 31 42 89 54 32 70 90 66 33 60 94 97 34 59 95 thirty 35 66 101 70 36 68 102 38 37 60 103 79 38 52

Claims (90)

CLAIM 1. The use of a compound for the preparation of a medicament for the treatment of cancer, characterized in that the compound is selected from or their pharmaceutically acceptable salts. 2. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 3. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. - 72 040967 4. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 5. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 6. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 7. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 8. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 9. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 10. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 11. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. - 73 040967 12. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 13. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 14. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 15. Use according to claim 1, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 16. Use according to claim 1, characterized in that the compound is 17. Use according to claim 1, characterized in that the compound is OH OH 18. Use according to claim 1, characterized in that the compound is 19. Use according to claim 1, characterized in that the compound is - 74 040967 20. Application according to 1, characterized in that the connection is 21. Application according to 1, characterized in that the connection is 1, characterized in that the connection is BUT. BUT. HE 22. Application according to 23. Application according to t — ό ^{n n} A. 1, characterized in that the connection is 24. Application according to 1, characterized in that the connection is 25. Application according to 1, characterized in that the connection is 26. Application according to 1, characterized in that the connection is 1, characterized in that the connection is 27. Application according to 28. Application according to 1, characterized in that the connection is - 75 040967 29. Use according to claim 1, characterized in that the compound is 30. Use according to any one of claims 1 to 29, characterized in that the cancer is selected from lung cancer, breast cancer, colon cancer, kidney cancer, bladder cancer, thyroid cancer, prostate cancer, osteosarcoma and Hodgkin's lymphoma. 31. The use of a compound for the treatment of cancer, characterized in that the compound is selected from or their pharmaceutically acceptable salts. 32. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. - 76 040967 33. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 34. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 35. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 36. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 37. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 38. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 39. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 40. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. - 77 040967 41. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 42. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 43. The use of a compound according to claim 31, characterized in that the compound is BUT HE H ₂ N N--O or a pharmaceutically acceptable salt thereof. 44. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 45. The use of a compound according to claim 31, characterized in that the compound is or a pharmaceutically acceptable salt thereof. 46. The use of a compound according to claim 31, characterized in that the compound is 47. The use of a compound according to claim 31, characterized in that the compound is 48. The use of a compound according to claim 31, characterized in that the compound is 49. The use of a compound according to claim 31, characterized in that the compound is - 78 040967 50. The use of a compound according to claim 31, characterized in that the compound is 51. The use of a compound according to claim 31, characterized in that the compound is 52. The use of a compound according to claim 31, characterized in that the compound is 53. The use of a compound according to claim 31, characterized in that the compound is 54. The use of a compound according to claim 31, characterized in that the compound is 55. The use of a compound according to claim 31, characterized in that the compound is 56. The use of a compound according to claim 31, characterized in that the compound is 57. The use of a compound according to claim 31, characterized in that the compound is - 79 040967 58. The use of a compound according to claim 31, characterized in that the compound is 59. The use of a compound according to claim 31, characterized in that the compound is 60. Use of a compound according to any one of claims 31 to 59, wherein the cancer is selected from lung cancer, breast cancer, colon cancer, kidney cancer, bladder cancer, thyroid cancer, prostate cancer, osteosarcoma and Hodgkin's lymphoma . 61. A method of treating cancer in a subject in need thereof, comprising administering to the subject a compound selected from or their pharmaceutically acceptable salts. - 80 040967 62. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 63. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 64. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 65. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 66. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 67. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 68. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 69. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. - 81 040967 70. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 71. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 72. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 73. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 74. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 75. The method of claim 61, wherein the compound is or a pharmaceutically acceptable salt thereof. 76. The method of claim 61, wherein the compound is 77. The method of claim 61, wherein the compound is - 82 040967 78. The method of claim 61, wherein the compound is 79. The method of claim 61, wherein the compound is 80. The method of claim 61, wherein the compound is 81. The method of claim 61, wherein the compound is 82. The method of claim 61, wherein the compound is 83. The method of claim 61, wherein the compound is 84. The method of claim 61, wherein the compound is 85. Way to 61, characterized in that the connection is 86. The method of claim 61, wherein the compound is - 83 040967 87. The method of claim 61, wherein the compound is 88. The method of claim 61, wherein the compound is 89. The method of claim 61, wherein the compound is 90. The method of any one of claims 61-89, wherein the cancer is selected from lung cancer, breast cancer, colon cancer, kidney cancer, bladder cancer, thyroid cancer, prostate cancer, osteosarcoma, and Hodgkin's lymphoma.