CN110300759A - The new compound of anti-mycobacteria - Google Patents

The new compound of anti-mycobacteria Download PDF

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Publication number
CN110300759A
CN110300759A CN201780086783.1A CN201780086783A CN110300759A CN 110300759 A CN110300759 A CN 110300759A CN 201780086783 A CN201780086783 A CN 201780086783A CN 110300759 A CN110300759 A CN 110300759A
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Prior art keywords
alkyl
compound
amino
bacterium
base
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CN201780086783.1A
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Chinese (zh)
Inventor
R·J·佩恩
W·J·布里顿
A·特兰
W·特兰
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University of Sydney
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University of Sydney
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Priority claimed from AU2016905229A external-priority patent/AU2016905229A0/en
Application filed by University of Sydney filed Critical University of Sydney
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/08—Antibacterial agents for leprosy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/06—Antibacterial agents for tuberculosis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0215—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0217—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing the structure -C(=O)-C-N-C(=O)-N-C-C(=O)-
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Abstract

The present invention relates to antimicrobial compounds.Particularly, these compounds can be used for inhibiting the growth of bacterium, especially mycobacterium tuberculosis (Mtb), and/or the bacterium with the targeting of phosphoric acid-N-MurNAc- pentapeptide translocase.The invention further relates to the application methods containing the composition of these compounds and these compounds and composition.

Description

The new compound of anti-mycobacteria
Technical field
The present invention relates to antimicrobial compounds.Particularly, the compounds of this invention is used to inhibit the growth of bacterium, especially tuberculosis Mycobacteria (Mtb), and/or targeting have phosphoric acid-N-MurNAc- pentapeptide translocase (phospho-MurNAc- Pentapeptide translocase) bacterium.The invention further relates to contain the composition of these compounds and these changes Close the application method of object and composition.
Background technique
Tuberculosis (TB) is as caused by bacterium mycobacterium tuberculosis (Mtb) infection.According to the data of the World Health Organization, Tuberculosis has resulted in 1,500,000 people death, and neopathy number of cases in 2014 is 9,600,000.
The six months by a definite date tetrads that the treatment of TB is Rimactazid, ethambutol and pyrazinamide are treated at present Method.The therapy provides the special cure rate more than 95% for drug susceptibility tuberculosis, however, it is to multidrug resistance (MDR) It is invalid with extensive drug resistance (XDR) TB.This is of increasing concern in the world.The two wires antibiotic of long-term use can be controlled effectively MDR infection is treated, but actually there are no the treatment methods that can be used for XDR infection.
Recently, drug shellfish is up to quinoline (bedaquiline) and Di Lamani (delamanid) respectively by U.S.'s food and medicine Management board (FDA) and European drug administration (EMA) ratify to be used for resistant tuberculosis (TB).However, since potential toxicity is asked Topic, these drugs only use as a last resort.
It is effective, hypotoxicity in order to provide drug susceptibility-types, multidrug resistance type and the treatment lungy of extensive drug-resistant type Selection, there is an urgent need to the novel tuberculosis drugs with new role mode.
In the present specification to the reference of any background technique all it is not an admission that or in any way implying the background technique Common knowledge is constituted in any other judicial region, or can reasonably expect that those skilled in the art can recognize the background technique Determine, understand or be considered as correlation herein, and/or combines other prior arts of this field.
Summary of the invention
This application involves a kind of compounds, especially antimicrobial compound.The biology of these compounds inhibition bacteria cell wall Synthesis and/or bacterial growth, and have shown that mycobacterium tuberculosis (Mtb) especially effectively.
On the one hand, the present invention provides Formulas I compound represented:
Or its salt, solvate, polymorph or prodrug；
Wherein,
For singly-bound or double bond；Wherein whenWhen for singly-bound, the spatial chemistry of the position is R- configuration.
R1Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, fragrant amino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R2Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R3For-C (O) R4Or-CH2R4；With
R4It is selected from the group: hydroxyl, or optionally replace: C1-C15Alkyl, C1-C15Alkyl amino；C1-C15Alkoxy；C1- C15Alkyl sulfenyl；C1-C15Halogenated alkyl, C1-C15Halogenated alkoxy, C1-C15Hydroxy alkyl, C1-C15Alkyl carboxyl, C1-C15Alkane Yl-carboxamides, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1-C4Alkyl) C3-C7Heterocycle, virtue Base, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1- C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
Collateral condition is:
WhenWhen for double bond, R3It is not-COOH, and
WhenFor singly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl ammonia Base ,-CH (CH3)OH、-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2S CH3；And R2For methyl, isopropyl Base, isobutyl group ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Replace Benzyl, aryl replace benzyl, naphthalene or-CH2When cyclohexyl；R3It is not-COOH.
On the other hand, the present invention provides Formula II compound:
Or its salt, solvate, polymorph or prodrug；
Wherein,
For singly-bound or double bond；Wherein, whenWhen for singly-bound, the spatial chemistry of the position is R- configuration.
R1Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, fragrant amino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R2Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R3For-C (O) R4Or-CH2R4；With
R4It is selected from the group: hydroxyl, or optionally replace: C1-C15Alkyl, C1-C15Alkyl amino；C1-C15Alkoxy；C1- C15Alkyl sulfenyl；C1-C15Halogenated alkyl, C1-C15Halogenated alkoxy, C1-C15Hydroxy alkyl, C1-C15Alkyl carboxyl, C1-C15Alkane Yl-carboxamides, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1-C4Alkyl) C3-C7Heterocycle, virtue Base, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1- C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
Collateral condition:
WhenWhen for double bond, R3It is not-COOH, and
WhenFor singly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl Amino ,-CH (CH3)OH、-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2SCH3；And R2For methyl, isopropyl Base, isobutyl group ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Replace Benzyl, aryl replace benzyl, naphthalene or-CH2When cyclohexyl；R3It is not-COOH.
On the other hand, the present invention provides a kind of pharmaceutical composition, it includes Formulas I and/or Formula II compound and pharmaceutically may be used The excipient of receiving.
On the other hand, the present invention provides a kind of method prevented and/or treat the disease or illness that are adjusted by bacterium, including Give a effective amount of Formulas I of mammalian therapeutic and/or Formula II compound or comprising Formulas I and/or Formula II compound of this needs With the composition of pharmaceutically acceptable excipient.
Bacterium can be Gram-positive or Gram-negative.
Therefore, on the one hand, the present invention provides a kind of disease prevented and/or treatment is adjusted by mycobacterium tuberculosis (Mtb) Or the method for illness comprising to have this need mammal give therapeutically effective amount Formulas I and/or Formula II compound or Composition comprising Formulas I and/or Formula II compound and pharmaceutically acceptable excipient.
On the other hand, the present invention provide it is a kind of prevention and/or treatment by with phosphoric acid-N-MurNAc- pentapeptide translocase The method of disease or illness that bacterium is adjusted, this method include giving a effective amount of present invention of mammalian therapeutic of this needs The compound of Formulas I and/or Formula II or composition comprising Formulas I and/or Formula II compound and pharmaceutically acceptable excipient.
On the one hand, the present invention provides a kind of prevention and/or treats method lungy comprising to the lactation for having this to need Animal gives the Formulas I and/or Formula II compound or comprising Formulas I and/or Formula II compound and pharmaceutically acceptable of therapeutically effective amount Excipient composition.
On the other hand, the present invention provides Formulas I and/or Formula II compound or comprising Formulas I and/or Formula II compound and pharmaceutically The composition of acceptable excipient is in preparation for preventing or treating in the disease or illness that are adjusted by gram-positive bacterium Purposes.
On the one hand, the present invention provides Formulas I and/or Formula II compound or comprising Formulas I and/or Formula II compound and pharmaceutically may be used The composition of the excipient of receiving is in preparation for preventing or treating by mycobacterium tuberculosis (Mtb) disease adjusted or illness Purposes in drug.
On the other hand, the present invention provides Formulas I and/or Formula II compound or comprising Formulas I and/or Formula II compound and pharmaceutically The composition of acceptable excipient is in preparation for preventing and/or treating by with phosphoric acid-N-MurNAc- pentapeptide translocase Purposes in the drug of disease or illness that bacterium is adjusted.
On the one hand, the present invention provides the compound of Formulas I and/or Formula II or compound and pharmacy comprising Formulas I and/or Formula II The composition of upper acceptable excipient is in preparation for preventing and/or treating the purposes in tuberculosis.
On the other hand, the present invention provides the compound of Formulas I and/or Formula II or compound and medicine comprising Formulas I and/or Formula II The composition of acceptable excipient on is used to prevent and/or treat by bacterium (such as: Gram-positive or gram-negative Property bacterium) regulation disease.
On the other hand, the present invention provides the compound of Formulas I and/or Formula II or compound and medicine comprising Formulas I and/or Formula II The composition of acceptable excipient on is used to prevent and/or treat the disease regulated and controled by mycobacterium tuberculosis (Mtb).
On the other hand, the present invention provides the compound of Formulas I and/or Formula II or compound and medicine comprising Formulas I and/or Formula II The composition of acceptable excipient on is used to prevent and/or treat by with phosphoric acid-N-MurNAc- pentapeptide translocase Bacterial regulatory disease.
On the one hand, the present invention provides the compound of Formulas I and/or Formula II or compound and pharmacy comprising Formulas I and/or Formula II The composition of upper acceptable excipient, is used to prevent and/or treat tuberculosis.
As used herein, unless the context otherwise requires, the variant of term " includes " and the term, such as "comprising", " packet Containing " and " by comprising ", it is no intended to exclude other additives, component, integer or step.
The further embodiment of aspect described in other aspects of the present invention and preceding paragraphs will be become by following description It is clear that these descriptions are by example and to be given with reference to the accompanying drawings.
Detailed description of the invention
Inhibiting effect of Fig. 1 analog 7 to MtbH37Rv.
Inhibiting effect of Fig. 2 analog 8 to MtbH37Rv.
Inhibiting effect of Fig. 3 analog 9 to MtbH37Rv.
Inhibiting effect of Fig. 4 analog 10 to MtbH37Rv.
Inhibiting effect of Fig. 5 analog 11 to MtbH37Rv.
Inhibiting effect of Fig. 6 analog 21 to MtbH37Rv.
Inhibiting effect of Fig. 7 analog 22 to MtbH37Rv.
Inhibiting effect of Fig. 8 analog 23 to MtbH37Rv.
Inhibiting effect of Fig. 9 analog 24 to MtbH37Rv.
Inhibiting effect of Figure 10 analog 25 to MtbH37Rv.
Inhibiting effect of Figure 11 analog 26 to MtbH37Rv.
Inhibiting effect of Figure 12 analog 27 to MtbH37Rv.
Inhibiting effect of Figure 13 analog 28 to MtbH37Rv.
Inhibiting effect of Figure 14 analog 30 to MtbH37Rv.
Inhibiting effect of Figure 15 analog 31 to MtbH37Rv.
Inhibiting effect of Figure 16 analog 32 to MtbH37Rv.
Inhibiting effect of Figure 17 analog 33 to MtbH37Rv.
Inhibiting effect of Figure 18 analog 34 to MtbH37Rv.
Inhibiting effect of Figure 19 analog 35 to MtbH37Rv.
Inhibiting effect of Figure 20 analog 36 to MtbH37Rv.
Inhibiting effect of Figure 21 analog 37 to MtbH37Rv.
Figure 22 is in THP-1 cell, inhibiting effect of the analog 25 to MtbH37Ra.IC50Represent the flat of 2 independent experiments Mean value, each experiment is in triplicate.
Figure 23 is in THP-1 cell, inhibiting effect of the analog 36 to Mtb H37Ra.IC50Represent 2 independent experiments Average value, each experiment is in triplicate.
Figure 24 is in THP-1 cell, inhibiting effect of the analog 37 to MtbH37Ra.IC50Represent the flat of 2 independent experiments Mean value, each experiment is in triplicate.
Figure 25 comes from Mtbmc2The TLC measurement of 6230 films inhibits MurX for analog 25.A:TLC；B: analog 25 presses down Original doses-response curve of MurX processed；C: the Logarithm conversion dose-response curve of the inhibition of analog 25 MurX.
Figure 26 comes from Mtbmc2The TLC measurement of 6230 films inhibits MurX for analog 36.A:TLC；B: analog 36 presses down Original doses-response curve of MurX processed；C: the Logarithm conversion dose-response curve of the inhibition of analog 36 MurX.
Figure 27 comes from Mtbmc2The TLC measurement of 6230 films inhibits MurX for analog 37.A:TLC；B: analog 37 presses down Original doses-response curve of MurX processed；C: the Logarithm conversion dose-response curve of the inhibition of analog 37 MurX.
Figure 28 comes from Mtbmc2The TLC measurement of 6230 films inhibits MurX for similar tunicamycin.A:TLC；B: tunicamycin Inhibit original doses-response curve of MurX；C: the Logarithm conversion dose-response curve of tunicamycin inhibition MurX.
The kinetic parameter of Figure 29 MurX is evaluated.A: Michaelis-Menten equation (Michaelis-Menten) figure；B: protoenzyme dynamics Data.Data represent the average value of independent experiment twice.
Figure 30 comes from Mtbmc2The fluoremetry of 6230 films inhibits MurX for analog 25.A: analog 25 inhibits Original doses-response curve of MurX；B: the Logarithm conversion dose-response curve of the inhibition of analog 25 MurX.The data of offer It is the average value of independent experiment twice.
Figure 31 comes from Mtbmc2The fluoremetry of 6230 films inhibits MurX for analog 36.A: analog 36 inhibits Original doses-response curve of MurX；B: the Logarithm conversion dose-response curve of the inhibition of analog 36 MurX.The data of offer It is the average value of independent experiment twice.
Figure 32 comes from Mtbmc2The fluoremetry of 6230 films inhibits MurX for analog 37.A: analog 37 inhibits Original doses-response curve of MurX；B: the Logarithm conversion dose-response curve of the inhibition of analog 37 MurX.The data of offer It is the average value of independent experiment twice.
Figure 33 comes from Mtbmc2The fluoremetry of 6230 films inhibits MurX (positive control) for tunicamycin.A: tunicamycin Inhibit original doses-response curve of MurX；B: the Logarithm conversion dose-response curve of tunicamycin inhibition MurX.The number of offer According to the average value for being independent experiment twice.
Inhibiting effect of Figure 34 analog 71 to MtbH37Rv.
Inhibiting effect of Figure 35 analog 79 to MtbH37Rv.
Figure 36 is in THP-1 cell, inhibiting effect of the analog 78 to MtbH37Rv.
Figure 37 is in THP-1 cell, inhibiting effect of the analog 79 to Mtb H37Rv.
Specific embodiment
The present invention describes the compound for preventing and/or treating the disease or illness that are adjusted by bacterium.According to this hair Bright, disease or illness can be related to Related Bacteria infection, are induced by it or are caused by it.Preferably, bacterium is gram sun Property bacterium.Even further preferably, bacterium is mycobacterium tuberculosis (Mtb).
On the one hand, compound provided herein can be used for preventing and/or treating the illness adjusted by bacterium.Be not intended to by appoint What theoretical constraint, it is assumed that the activity of these compounds is by inhibiting bacterial growth and/or inhibiting bacteria cell wall biosynthesis Come what is realized.The compound has special selectivity to mycobacterium tuberculosis (Mtb), therefore can be used for treating such as tuberculosis Disease.The compound can also be used to preventing and/or treating the other illnesss as caused by bacterium.
On the one hand, the present invention provides Formulas I compound represented:
Or its salt, solvate, polymorph or prodrug；
Wherein,
For singly-bound or double bond；Wherein whenWhen for singly-bound, the spatial chemistry of the position is R- configuration.
R1Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, fragrant amino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R2Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R3For-C (O) R4Or-CH2R4；With
R4It is selected from the group: hydroxyl, or optionally replace: C1-C15Alkyl, C1-C15Alkyl amino；C1-C15Alkoxy；C1- C15Alkyl sulfenyl；C1-C15Halogenated alkyl, C1-C15Halogenated alkoxy, C1-C15Hydroxy alkyl, C1-C15Alkyl carboxyl, C1-C15Alkane Yl-carboxamides, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1-C4Alkyl) C3-C7Heterocycle, virtue Base, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1- C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
Collateral condition:
WhenWhen for double bond, R3It is not-COOH, and
WhenFor singly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl Amino ,-CH (CH3)OH、-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2SCH3；And R2For methyl, isopropyl Base, isobutyl group ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Replace Benzyl, aryl replace benzyl, naphthalene or-CH2When cyclohexyl；R3It is not-COOH.
On the one hand, the present invention provides Formula II compound:
Or its salt, solvate, polymorph or prodrug；
Wherein,
For singly-bound or double bond；Wherein whenWhen for singly-bound, the spatial chemistry of the position is R- configuration.
R1Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, fragrant amino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R2Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkane Base, C1-C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxyl Base alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle Base；(C1-C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and fragrant acyl Base；
R3For-C (O) R4Or-CH2R4；With
R4It is selected from the group: hydroxyl, or optionally replace: C1-C15Alkyl, C1-C15Alkyl amino；C1-C15Alkoxy；C1- C15Alkyl sulfenyl；C1-C15Halogenated alkyl, C1-C15Halogenated alkoxy, C1-C15Hydroxy alkyl, C1-C15Alkyl carboxyl, C1-C15Alkane Yl-carboxamides, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1-C4Alkyl) C3-C7Heterocycle, virtue Base, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1- C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
Collateral condition:
WhenWhen for double bond, R3It is not-COOH, and
WhenFor singly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl Amino ,-CH (CH3)OH、-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2SCH3；And R2For methyl, isopropyl Base, isobutyl group ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Replace Benzyl, aryl replace benzyl, naphthalene or-CH2When cyclohexyl；R3It is not-COOH.
In one embodiment, which is formula III compound represented:
Or its salt, solvate, polymorph or prodrug；
Wherein R1, R2And R4Definition as described in Formula II,
Collateral condition:
WhenWhen for double bond, R4It is not-OH, and
WhenFor singly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl ammonia Base ,-CH (CH3) OH ,-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2SCH3；And R2For methyl, isopropyl Base, isobutyl group ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Replace Benzyl, aryl replace benzyl, naphthalene or-CH2When cyclohexyl；R4It is not-OH.
In one embodiment, compound is formula IV compound represented:
Or its salt, solvate, polymorph or prodrug；
Wherein, R1, R2And R4Definition as described in Formula II.
Formulas I, II, the preferred embodiment of III and/or IV compound are described below.
Preferably,For singly-bound.
Preferably, R1It is selected from the group: hydrogen, or the following group optionally replaced: C1-C6Alkyl, C1-C2Aralkyl optionally replaces (C1-C2Alkyl) heteroaryl, C1-C6Alkyl amino；C1-C6Alkoxy, C1-C6Alkyl carboxyl, C1-C6Hydroxy alkyl.It is even more excellent Selection of land, R1It is selected from the group: benzyl, the naphthalene, C that hydrogen, methyl, hydroxyl replace2-C4Alkyl amino, cyclohexyl ,-CH2Cyclohexyl Or-CH (OH) CH3。
Preferably, R2Selected from the following group comprising optionally replacing: C1-C6Alkyl, (C1-C2)C3-C7Naphthenic base, C1-C4Aralkyl And C1-C6Alkyl sulfenyl.Even further preferably, R2It is selected from the group: isopropyl ,-CH2Naphthalene ,-CH2Cyclohexyl ,- CH2CH2SCH3；-CH2CH2S(O)CH3With
Preferably, R4Selected from the group comprising optionally replacing the following group: C1-C15Alkoxy, C1-C4Aralkyl；C1-C15Alkyl Amino；C1-C4Alkoxy, C1-C4Aryl alkyl amino and C1-C15Alkyl amino.It is highly preferred that R4It is selected from the group: methoxyl group, own oxygen Base, dodecyloxy, hydroxyl ,-CH2C(CH3)3,-O- benzyl ,-NH- benzyl ,-NH- benzyl, hexylamino.Even more preferably Ground, R3It is selected from the group :-CH2C(CH3)3,-O- benzyl ,-NH- benzyl and own amino.
In some preferred embodiments, compound compound selected from the group below:
Or its salt, solvate, polymorph or prodrug.
In one embodiment, the compound of Formulas I and/or Formula II is selected from the compound comprising the following group:
Or its salt, solvate, polymorph or prodrug.
In a preferred embodiment, the compound of Formulas I and/or Formula II is selected from the compound comprising the following group:
On the one hand, the present invention provides compound selected from the following:
Preferably, compound is
Definition
As used herein, term " alkyl " refers to saturations or undersaturated straight chain or branched hydrocarbyl, has 1 to 15 carbon Atom or intervenient any range, i.e., it includes 1,2,3,4,5,6,7,8,9,10,11,12,13,14 or 15 carbon originals Son.Therefore, which includes saturated alkyl and alkenyl and alkynyl.Alkyl is optionally substituted with a substituent, and allows multiple degree of substitution. The example of " alkyl " used herein includes but is not limited to methyl, ethyl, n-propyl, isopropyl, acrylic, the positive fourth of propinyl Base, cyclobutenyl, isobutyl group, tert-butyl, n-pentyl, pentenyl, isopentyl etc..Saturated hydrocarbyl can be mono-, double- or multi- insatiable hunger Sum, and double bond (alkenyl) or three (alkynyl) keys can be contained.
As used herein, term " C1-C3Alkyl ", " C1-C4Alkyl ", " C1-C6Alkyl " and " C1-C15Alkyl " refers to as above Definition contain at least one, be respectively provided with the alkyl of at most 3,4,6 or 15 carbon atoms, or therebetween any range (for example, Alkyl containing 2-5 carbon atom is also in C1-C6In the range of).
As used herein, term " halogen " refers to fluorine (F), chlorine (Cl), and bromine (Br) or iodine (I), term " halogenated " refer to halogen Plain radical fluoro (- F), chloro (- Cl), bromo (- Br) and iodo (- I).Preferably, ' halogenated ' is fluoro or chloro.
As used herein, term " naphthenic base " refers to non-aromatic hydrocarbon ring.In a similar way, term " C3-C7Naphthenic base " is Refer to the non-aromatic naphthenic ring with 3-7 carbon atom or any range therebetween.For example, C3-C7Naphthenic base further includes containing 4-6 The naphthenic base of a carbon atom.Alkyl is as defined above, and can be substituted.Exemplary " C for use in the present invention3-C7 Naphthenic base " includes but is not limited to cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl and suberyl.This ring optionally with one or Multiple naphthenic base that other optionally replace, " heterocycle ", aromatic ring or hetero-aromatic ring are condensed.
As used herein, term " heterocycle " or " heterocycle " refer to non-aromatic heterocyclic, for saturation or have one or Multiple degrees of unsaturation are selected from S, S (O), S (O) containing one or more hetero atoms2, the substitution of O or N.Term " C3-C7Heterocycle Base " refers to the non-aromatic naphthenic ring with three to seven carbon atoms, contains one or more hetero atoms as described herein and takes Generation.Heterocyclic moiety can be substituted, and allow multiple degree of substitution.Term " C3-C7Heterocycle " further includes containing C4-C5, C5-C7, C6- C7, C4-C7, C4-C6And C5-C6The heterocycle of carbon atom.Preferably, heterocycle contains 4-6 carbon atom and one or two miscellaneous original Son.It is highly preferred that heterocycle contains, there are five carbon atoms and a hetero atom or four carbon atom and two hetero atoms or five carbon Atom and a hetero atom.This ring optionally with one or more " heterocycles " that other optionally replace, naphthenic base, aryl or It is heteroaryl-condensed.The example of heterocycle includes but is not limited to tetrahydrofuran, pyrans, oxetanes, 1,4- dioxanes, 1,3- bis- Oxane, piperidines, piperazine, N methyl piperazine base, 2,4- piperazinedione, pyrrolidines, imidazolidine, pyrazolidine, morpholine, thiomorpholine, Tetrahydric thiapyran, thiophane etc..The heterocyclic system of a substitution of the invention be the coumarin ring of substitution, such as 6,7- dimethoxy Butylcoumariii.
Term " (C1-C2Alkyl) C3-C7Heterocycle " includes the alkyl that contains 1 or 2 carbon atom as compound and miscellaneous Ring (i.e. heterocycle ,-CH2Heterocycle or-CH2CH2Heterocycle) between linker.These heterocycles can be further substituted.
Substituted naphthenic base and heterocycle can be replaced by any suitable substituent group as described below.
As used herein, term " aryl " refers to the phenyl ring optionally replaced or the benzene ring system optionally replaced and one or more A phenyl ring optionally replaced is condensed, to form such as anthracene, phenanthrene or naphthalene ring system.The example of " aryl " include but is not limited to phenyl, 2- naphthalene, 1- naphthalene, xenyl and its derivative replaced.
As used herein, term " heteroaryl " refers to five yuan, hexa-atomic or seven unit monocycle aromatic rings, or refer to comprising at least one five Member, hexa-atomic or seven unit monocycle aromatic rings condensed-bicyclics or tricyclic aromatic ring system.These hetero-aromatic rings contain one or more nitrogen, sulphur And/or oxygen heteroatom, wherein N- oxide and oxysulfide and dioxide are allowed hetero atom substitution, and can be optional Ground is replaced by most three atoms.The example of " heteroaryl " group used herein includes furyl, thienyl, pyrrole radicals, miaow Oxazolyl, pyrazolyl, triazolyl, tetrazole radical, thiazolyl, oxazolyl, isoxazolyl, oxadiazoles base, oxo pyridine base, thiadiazoles Base, isothiazolyl, pyridyl group, pyridazinyl, pyrazinyl, pyrimidine radicals, quinolyl, isoquinolyl, benzofuranyl, benzothiophene Base, indyl, indazolyl, benzimidazolyl and its substitution form.
As used herein, term " alkyl sulfenyl " refers to alkyl as defined above, former by one or more sulphur (- S-) Son or methylthio group bonding, or in alkyl chain end or one or more sulphur atoms or group are contained in end.It includes, for example, Following groups: mercaptan, thioesters, thioether, alkyl sulfur compounds, alkyl sulphonyl, alkyl sulfoxide base, alkyl sulfuryl, alkyl sulfenic acids, Alkyl sulfinic acid and chain alkyl sulfonic acid.Some non-limiting examples of alkyl sulfenyl include-CH2CH2SO2CH3、-CH2S(O)OH、- SO2CH2CH3、-CH2S-OH、-CH2SH、-CH3CH2SCH3、CH2CH2S(O)CH3、CH2CH2SO3H.As described below, these groups can To be further substituted, to provide the group of such as alkyl sulfonyl amino.
As used herein, term " alkoxy " refers to alkyl as defined above, passes through one or more oxygen (- O-) atoms Bonding, or contain one or more oxygen atoms in alkyl chain.Therefore, these groups include ester, ether, ketone and aldehyde.Alkoxy Some non-limiting examples include-CH2CH2OCH3、-OCH2CH3、-CO2CH2CH3、-CH2OH、-CH3CH2C(O)CH3、- CH2CH2COOCH3、-OCH2CH2OCH3Deng.As described below, these groups can be further substituted.
As used herein, term " alkyl amino " refers to alkyl chain as defined above comprising the amido in alkyl chain, Or the amine of the section start in the chain adjacent with the substituent group of the rest part of compound, or in the end of chain.For example, alkyl amino It can be following group :-CH2CH2NHCH3、-NHCH2CH3、-CH2N(CH3)2、-CH2CH2NH2Or the like.Depending on substituent group In conjunction with position, such as amide is (for example, wherein substituent group is in R3Place passes through amine combination).As described below, alkyl amino can be into One step is substituted.
As used herein, term " halogenated alkyl " refers to the alkyl as defined above being bonded with halogen group.Halogenated alkyl Some examples include-CH2F、-CF3With-CH2CH2CH2CHCl2.In the conceived case, these groups can further be taken Generation, as described below.For example,-CH2C(O)CH2CHCl2。
As used herein, term " hydroxy alkyl " refers to the alkyl as defined above being bonded with hydroxyl.As described below, these Group can be further substituted.
As used herein, term " alkyl carboxyl " refers to the alkyl as defined above with carboxylic-bond.As described below, these Group can be further substituted.
As used herein, term " halogenated alkoxy " refers to the alkoxy as defined above being bonded with halogen group.It is as follows Described, these groups can be further substituted.
As used herein, term " alkylcarboxamide " refers to the alkyl as defined above being bonded with carboxylacyl amine group.It is as follows Described, these groups can be further substituted.
As used herein, term " aryloxy group ", " arylamino " or " arylthio " refers to passes through oxygen (- O-) atom respectively, Amino or the aryl as defined above of sulfenyl bonding.Amino and thio group can be further substituted, as described below.
As used herein, term " aralkyl " refers to through aryl alkyl linked as defined above.As described below, these Group can be further substituted.When the group has specific carbon atom range, such as " C1-C4Aralkyl ", C1-C4It is Refer to the carbon atom number in the hydrocarbyl component of the group.
As used herein, term " aralkoxy " refers to the aryl being bonded by alkoxy as defined above.As described below, These groups can be further substituted.When the group has specific carbon atom range, such as " C1-C4Aralkoxy ", C1-C4Refer to the carbon atom number in the alkyl component of the group.
As used herein, term " aryl alkyl amino " refers to the aryl being bonded by alkyl amino as defined above.It is as follows Described, these groups can be further substituted.When the group has specific carbon atom range, such as " C1-C4Aralkyl Amino ", C1-C4Refer to the carbon atom number in the alkyl component of the group.
" substituent group " refers to the molecular moiety being bonded with the atom covalence in target molecule as used herein.For example, " ring Substituent group " can be such as halogen, the part of alkyl or other substituent groups as described herein, and the atom as ring members, It is preferred that carbon or nitrogen-atoms covalent bonding.As used herein, term " substituted " refers to any one or more on specified atom Hydrogen is replaced selected from specified substituent group, and condition is no more than the common fare of specified atom, and replaces the compound of generation It is stable, it can separation, characterization and the compound for carrying out biological activity test.
Throughout the specification, the term " optionally replacing " used or " can be substituted " etc. indicate that the group can be with It is further substituted with or without one or more non-hydrogen substituent groups or condensed (form polycyclic system).For suitable chemistry The substituent group of upper suitable particular functional group it will be apparent to those skilled in the art that.
The example of substituent group includes but is not limited to:
C1-C6Alkyl, C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxy alkyl, C3-C7Heterocycle, C3-C7Ring Alkyl, C1-C6Alkoxy, C1-C6Alkyl sulfenyl, C1-C6Alkyl carboxyl, alkylamide, oxo, hydroxyl, thio, amino, acyl group, Carboxyl, carbamoyl, aryl, aryloxy group, heteroaryl, amino-sulfonyl, aroyl, aroylamino, 4-hetaroylpyrazol, acyl-oxygen Base, aryl acyloxy, heteroaryl acyloxy, alkoxy carbonyl, nitro, cyano, halogen, urea groups or C1-C6Perfluoroalkyl.
In appropriate circumstances, any of these groups can further be replaced by any of above group.For example, alkyl ammonia Base or dialkyl amido, C1-C6Alkoxy etc..
The salt of the compound is preferably pharmaceutically acceptable, it should be appreciated that acceptable salt also belongs to this in non-pharmaceutical The range of invention, because they can be used as preparing the intermediate of pharmaceutically acceptable salt.
Term " pharmaceutically acceptable derivates " may include any pharmaceutically acceptable salt, hydrate or prodrug or appoint What his compound, or (direct or indirect) Formulas I, Formula II, the change of formula III and/or formula IV are capable of providing when giving subject Close object or its active metabolite or residue.
Suitable pharmaceutically acceptable salt includes but is not limited to the salt of pharmaceutically acceptable inorganic acid, such as: hydrochloric acid, The salt of sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfamic acid and hydrobromic acid or pharmaceutically acceptable organic acid, such as: second Acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, malic acid, citric acid, lactic acid, mucus, glucose Acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzene sulfonic acid, salicylic acid, p-aminobenzene sulfonic acid, asparagus fern Propylhomoserin, glutamic acid, edetic acid(EDTA), stearic acid, palmitinic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid and valeric acid.
Alkali salt include but is not limited to it is pharmaceutically acceptable cation formed those of, such as sodium, potassium, lithium, calcium, magnesium, Zinc, ammonium, alkylammonium (such as the salt formed by triethylamine), alkoxy ammonium (such as the salt formed with ethanol amine and ethylenediamine), gallbladder Alkali or amino acid (such as arginine, lysine or histidine) formation.Type and its formation about pharmaceutically acceptable salt General information is known to the skilled in the art, and as described in common article, for example, P.H.Stahl and C.G.Wermuth " pharmaceutical salts handbook " (Handbook of Pharmaceutical salt, the 1st edition, 2002, Wiley- VCH)。
The progress of nitrogenous base available reagent is quaternized, such as: elementary alkyl halide reagent: methyl, ethyl, propyl and fourth Base chloride, bromide and iodide；Dialkyl sulfate: dimethyl and dithyl sulfate；With other reagents.
Prodrug includes: amino acid residue compound, or with free amine group and Formulas I, Formula II, the chemical combination of formula III and/or formula IV The polypeptide chain of two or more (for example, two, three or four) amino acid residues that the acylamino- of object is covalently attached.Amino Sour residue includes the 20 kinds of naturally occurring amino acid usually indicated by three letter characters, further includes: 4- hydroxy-proline, hydroxyl Lysine, desmosine, isodensmosine, 3-Methyl histidine, norvaline, Beta-alanine, γ-aminobutyric acid, citrulling, high half Guang Propylhomoserin, homoserine, ornithine and methionine sulfone.Prodrug further include: carbonic ester, carbamate, amide and Arrcostab chemical combination Object is covalently bound to Formulas I by carbonyl carbon prodrug sidechain, Formula II, on the above-mentioned substituent group of formula III and/or formula IV.
Term " polymorph " includes Formulas I, Formula II, any crystal form of formula III and/or formula IV compound, such as nothing Water form, aqueous form, solvate forms and mixed solvent compound form.
" solvate " refers to the compound that the interaction of the compounds of this invention and solvent is formed.
The therapeutical uses of compound
Surprisingly, the inventors discovered that the compound of Formulas I, Formula II, formula III and/or formula IV has shown Mtb Imitate and/or selective inhibitory activity.These compounds destroy mycobacterium tuberculosis phosphoric acid-N-MurNAc- pentapeptide translocase The activity of (MurX or translocase I) is for lipid I (key intermediate of mycobacteria peptide glycan synthesis) biosynthesis Complete film enzyme.
Therefore, these compounds can be used for preventing and/or treating by mycobacterium tuberculosis (Mtb) and/or have enzyme phosphoric acid- The disease or illness that the bacterium of N-MurNAc- pentapeptide translocase (MurX) is adjusted.Therefore, the compound can be used for prevent and/or Treat tuberculosis.
The compound can also be used in other diseases caused by treating other bacteriums or illness, such as: it is Acinetobacter bauamnnii, big Enterobacteria, staphylococcus aureus, methicillin-resistant staphylococcus aureus, comma bacillus, produces gas intestines bar at pseudomonas aeruginosa Bacterium, produces alkali Providence, bacillus subtilis, enterococcus faecium, Yi Shi Li Site bacterium, epidermis grape ball at human pallid bacillus Bacterium, salmonella typhimurium and pseudoconcretion yersinia enterocolitica.
Preferably, other bacteriums are gram-positive.Preferably, these bacteriums are mycobacterium avium, leprosy branch bar Bacterium, mycobacterium abscessus, Mycobacterium bovis, bacillus subtilis, staphylococcus epidermis, enterococcus faecium, Yi Shi Li Site bacterium, gold Staphylococcus aureus, methicillin-resistant staphylococcus aureus.Even further preferably, these bacteriums are mycobacterium avium, leprosy Mycobacteria, mycobacterium abscessus and Mycobacterium bovis.
Therefore, which can be used for treating HIV/AIDS and cystic fibrosis, the machine in leprosy and bronchiectasis Opportunistic infections and the other diseases as caused by these or similar bacterium.
Therefore, on the one hand, the present invention provides a kind of pharmaceutical composition, and it includes Formulas I, Formula II, formula III and/or formula IV institutes Compound or its salt, solvate, polymorph or the prodrug and pharmaceutically acceptable excipient shown.
On the other hand, the present invention provides the method for disease or illness that prevention and/or treatment are adjusted by bacterium, including gives A effective amount of Formulas I of mammalian therapeutic for having this to need, Formula II are formula III and/or formula IV compound or its salt, solvate, more Crystal form object or prodrug, or include Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, polymorph or preceding The composition of medicine and pharmaceutically acceptable excipient.
In one embodiment, bacterium is Acinetobacter bauamnnii.In another embodiment, bacterium is large intestine bar Bacterium.In another embodiment, bacterium is pseudomonas aeruginosa.In another embodiment, bacterium is golden yellow grape Coccus.In another embodiment, bacterium is methicillin-resistant staphylococcus aureus.In another embodiment, carefully Bacterium is comma bacillus.In a further embodiment, bacterium is clostridium perfringen.In another embodiment, bacterium is Human pallid bacillus.In a further embodiment, bacterium is to produce alkali Providence.In another embodiment, carefully Bacterium is bacillus subtilis.In another embodiment, bacterium is enterococcus faecium.In a further embodiment, bacterium For Yi Shi Li Site bacterium.In another embodiment, bacterium is staphylococcus epidermis.In yet another embodiment, bacterium For salmonella typhimurium.In another embodiment, bacterium is pseudoconcretion yersinia enterocolitica.
Preferably, bacterium is gram-positive bacteria.Preferably, bacterium be mycobacterium tuberculosis (Mtb), mycobacterium avium, Mycobacterium leprae, mycobacterium abscessus, Mycobacterium bovis, bacillus subtilis, staphylococcus epidermis, enterococcus faecium, Yi Shi Li Site bacterium, staphylococcus aureus, methicillin-resistant staphylococcus aureus.Even further preferably, bacterium is tuberculosis branch Bacillus (Mtb), mycobacterium avium, Mycobacterium leprae, mycobacterium abscessus and Mycobacterium bovis.Most preferably, bacterium is knot Core mycobacteria (Mtb).
On the other hand, the present invention provides the method for disease or illness that prevention and/or treatment are adjusted by Mycobacterium tuberculosis, A effective amount of Formulas I of mammalian therapeutic including giving this needs, Formula II are formula III and/or formula IV compound or its salt, molten Agent compound, polymorph or prodrug, or include Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, polycrystalline The composition of type object or prodrug and pharmaceutically acceptable excipient.
On the other hand, the present invention provides prevention and/or treats by the bacterium with phosphoric acid-N-MurNAc- pentapeptide translocase The method of the disease or illness of adjusting, this method include giving a effective amount of formula I of mammalian therapeutic of this needs, Compound or its salt, solvate, polymorph or the prodrug of Formula II, formula III and/or formula IV, or include Formulas I, Formula II, formula The compound or salt of III and/or formula IV, the composition of solvate, polymorph or its prodrug and pharmaceutically acceptable tax Shape agent.
On the other hand, the present invention provides the method for disease or illness that prevention and/or treatment are adjusted by bacterium, including gives A effective amount of Formulas I of mammalian therapeutic for having this to need, Formula II are formula III and/or formula IV compound or its salt, solvate, more Crystal form object or prodrug, or include Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, polymorph or preceding The composition of medicine and pharmaceutically acceptable excipient.
On the other hand, the present invention provides Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, solvate, more Crystal form object or prodrug, or include Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, polymorph or preceding The composition of medicine and pharmaceutically acceptable excipient, is used to prepare prevention and/or treatment is adjusted by gram-positive bacterium Purposes in the drug of disease or illness.
On the other hand, the present invention provides Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, solvate, more Crystal form object or prodrug, or include Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, polymorph or preceding The composition of medicine and pharmaceutically acceptable excipient, is used to prepare prevention and/or treatment is adjusted by mycobacterium tuberculosis (Mtb) Purposes in the disease of section or the drug of illness.
On the other hand, the present invention provides Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, solvate, more Crystal form object or prodrug, or include Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, polymorph or preceding The composition of medicine and pharmaceutically acceptable excipient is used to prepare prevention and/or treatment by with phosphoric acid-N-MurNAc- five Purposes in the drug of the disease of the bacterial regulatory of peptide translocase.
On the other hand, the present invention provides Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, more Crystal form object or prodrug include Formulas I, Formula II, formula III and/or formula IV compound or its salt, solvate, polymorph or prodrug With the composition of pharmaceutically acceptable excipient, prepare for preventing and/or treating the purposes in tuberculosis.
On the other hand, the present invention provides Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, solvate, more The compound or its salt of crystal form object or prodrug, or comprising Formulas I, Formula II, formula III and/or formula IV, solvate, polymorph or The composition of prodrug and pharmaceutically acceptable excipient, it is excellent for preventing and/or treating the disease or illness that are adjusted by bacterium Select gram-positive bacterium.
On the other hand, the present invention provides Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, solvate, more The compound or its salt of crystal form object or prodrug, or comprising Formulas I, Formula II, formula III and/or formula IV, solvate, polymorph or The composition of prodrug and pharmaceutically acceptable excipient, for preventing and/or treating by mycobacterium tuberculosis (Mtb) adjusting Disease or illness.
On the other hand, the present invention provides Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, solvate, more The compound or its salt of crystal form object or prodrug, or comprising Formulas I, Formula II, formula III and/or formula IV, solvate, polymorph or The composition of prodrug and pharmaceutically acceptable excipient, for preventing and/or treating by with phosphoric acid-N-MurNAc- pentapeptide The disease or illness of the bacterial regulatory of translocase.
On the other hand, the present invention provides Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, solvate, more The compound or its salt of crystal form object or prodrug, or comprising Formulas I, Formula II, formula III and/or formula IV, solvate, polymorph or The composition of prodrug and pharmaceutically acceptable excipient is for preventing and/or treating tuberculosis.
Phosphoric acid-N-MurNAc- pentapeptide translocase can be MurX or MraY.Preferably, phosphoric acid-N-MurNAc- pentapeptide turns Position enzyme is MurX.
In one embodiment of the invention, in the above method and on the way, prevention or the tuberculosis (TB) treated are Multi-drug resistance tuberculosis (MDR TB).In another embodiment, in the above method and on the way, the tuberculosis of prevention or treatment It is extensive resistant tuberculosis (XDR TB).
In one embodiment, treatment of the TB to Rimactazid, ethambutol and/or pyrazinamide It is resistant.
In one aspect, mycobacterium tuberculosis (Mtb) is the H37Rv bacterial strain of Mtb.
Administration
Pharmaceutical composition can be by Formulas I, Formula II, compound or its salt, solvate, the polymorphic of formula III and/or formula IV Object or prodrug are formulated for any suitable administration route, including for example: local (for example, transdermal or eye), oral, oral cavity, Nasal cavity, vagina, rectum or parenteral administration.The term as used herein parenteral includes: subcutaneous, intradermal, intravascular (for example, quiet In arteries and veins), intramuscular, backbone, encephalic, intrathecal, intraocular, eye circumference, in socket of the eye, intrasynovial and intraperitoneal injection and any similar Injection or infusion techniques.In some embodiments it is preferred that being suitable for the composition of oral and extra-parenteral administration.For example, suitable Oral form includes: tablet, lozenge, pastille, aqueous or oily suspensions, dispersible powder or particle, lotion, hard or flexible glue Capsule or syrup or elixir.It, can be by one or more compounds and nothing for intravenous, intramuscular, subcutaneous or Intraperitoneal medication The mixing of bacterium aqueous solution, the aseptic aqueous solution are preferably isotonic with the blood of recipient.Said preparation can by by solid active at Divide and be dissolved in the water containing PHYSIOLOGICALLY COMPATIBLE substance (such as sodium chloride or glycine), there is the buffer pH compatible with physiological condition To generate aqueous solution, and keep the solution sterile to prepare.Preparation can exist in the form of unit or multi-dose container, such as seal Ampoule or bottle.The example of component is described in " Martindale-The Extra Pharmacopoeia " (Ma room temperature indale-The Extra Pharmacopoeia) " pharmaceutical science " in (Pharmaceutical Press, Britain, London 1993) and Ma room temperature in (ed.), Remington.
In the context of the present specification, the variant of term " administration " and the term includes " giving " and " administration ", including By any mode appropriate the compound of the present invention or composition are contacted, apply, deliver or provided to organism or surface.
For inhibiting Mtb and/or Mtb phosphoric acid-N-MurNAc- pentapeptide translocase (MurX), such as the H37Rv bacterial strain of Mtb, And other diseases and illness for preventing and/or treating tuberculosis and Mtb and the adjusting of other bacteriums, biology of the invention are living Property compound dosage can change in a wide range, can according to individual requirement be adjusted.Reactive compound of the invention Usually it is administered with therapeutically effective amount.Preferred dosage range is about 0.1mg to about 140mg per kg body weight per day (for example, one Patient about 0.5mg to about 7g) daily.Daily dose can be with single dose or multiple dose administration.It can be combined to produce with carrier material The amount of the active constituent of single formulation will change according to the subject and specific administration mode that are treated.Dosage unit form About 1mg is usually contained to about 500mg active constituent.
However, it should be understood that the specific dosage level of any particular subject will depend on many factors, comprising: spy used Determine activity, age, weight, general health, gender, diet, administration time, the administration route, discharge rate, medicine of compound Object combination (i.e. for treating the other drugs of subject), and receive the severity of the specified disease for the treatment of.If chemical combination Object is local administration rather than Formulations for systemic administration, and for preventing for treating, then dosage is usually lower.These treatments It can according to need and often carry out, and be judged as in the necessary time in treating physician and carry out.Those skilled in the art will manage Solution, Formulas I to be administered, Formula II, the dosage or therapeutically effective amount of formula III and/or formula IV compound may be needed for every Individual optimizes.The active constituent that pharmaceutical composition contains about 0.1 to 2000mg, preferably from about 0.5 to 500mg, most preferably About 1 to 200mg.Daily dose is about 0.01 to 100mg/kg weight, preferably from about 0.1 to about 50mg/kg weight, it may be possible to suitable 's.Daily dosage can be administered according to daily one to four dosage.Preferably, by daily single.
Term " therapeutically effective amount " or " effective quantity " refer to Formulas I, Formula II, the compound or its salt of formula III and/or formula IV, Solvate, polymorph or prodrug can improve or repair by the symptom of gram-positive bacterium (amount) adjusting disease Amount.
Term " treatment ", " treatment " and " treatment " herein for referring to curative therapy, prophylactic treatment and preventative Treatment.Therefore, in the context of the disclosure, term " treatment " includes curing, and improves or mitigates by gram-positive bacterium (such as Tuberculosis) adjust illness severity.
" prevention " or " prevention " refers to if after the compound of the present invention or pharmaceutical composition administration, prevents blue by leather The generation for the illness that family name's positive bacteria (such as tuberculosis) is adjusted, or mitigate the severity of illness.
As described above, the compounds of this invention can be administered together with pharmaceutical carrier, diluent or excipient.
Biological test
Using resazurin measuring method, the compounds of this invention to the H37Rv strain of Mtb is screened, it is then thin for HEK293 Born of the same parents carry out counter-selection choosing to measure the selectivity for being directed to one group of 15 kinds of Pathogenic gram-negatives and Gram positive bacteria strain.
When the compound of test is up to 200 μM of concentration, there is no cytotoxicity to HEK293 cell.In addition, most of tests Compound has selectivity to Mtb.
In addition, having carried out preliminary screening, also to assess compound to Mtb MurX inhibitory activity.This is related to Mtb mc2 The generation of 6230 memebrane protein preparations contains MurX and other memebrane proteins.By be added 200nM concentration given compound with And UDP- [14C] GlcNAc and UDP-MurNAc pentapeptide (Park's nucleotide) assesses the inhibiting effect for MurX.In enzymatic After the quenching of reaction and post-processing based on extraction, the inhibition journey of enzyme is measured using thin-layer chromatography (TLC) and Phosphorescence imaging Degree.The further details of these steps will discuss in following embodiment part.
Then these compounds are had studied to polyisopreneyl phosphoric acid-N-acetyl-glucosamine -1- phosphotransferase (WecA) inhibiting effect.However, the compounds of this invention of test does not have inhibiting effect to WecA, show that these compounds are peptides The selective depressant that lipid I is formed in glycan biosynthesis.
The measurement based on TLC is carried out using a series of several compounds of concentration, to determine IC50Value.It is good out as the result is shown Good activity, and show the IC for MtbMurX50There is between external activity significant correlation with MtbH37Rv.
The anti-mycobacteria activity of these compounds, institute are also assessed in the intracellular measurement using THP-1 macrophage It states THP-1 macrophage to be infected with Mtb, and measures the suppression of mycobacterium growth in the presence of a series of compound of concentration System.The compound of test maintains anti-mycobacteria activity to resist mycobacterium intracellulare growth.
Finally, testing stability of these compounds in mouse and human plasma and mouse and people's hepatomicrosome.Every kind Compound shows excellent stability, for people and mice plasma, degradation half life > 7h, and for people and Mouse Liver Microsomes, Reduce half-life period > 160 minute.
The compound tested shows the enzyme MurX for effectively inhibiting to be responsible for lipid I synthesis, this is in Mtb in the way of peptide glycan Key intermediate.The compound has selective active to Mtb.This selectivity be these compounds as TB drug and MurX provides practical advantage as the potential use of TB drug targets.
The further details of biological test can be seen in embodiment part.
It should be appreciated that in the present specification, disclosure of the invention and limiting is extended to mentioned above or from text Or in attached drawing two or more obvious independent features all optional combinations.All these different combinations constitute Various alternative aspects of the invention.
Method described herein and compound are described by following illustrative and non-limiting embodiment.
Embodiment
1. material and method
It is thin that analytic type is carried out in the silica plate (60 0.25mm F254 of Merck Kieselgel) of business preparation Layer chromatography (TLC).Flash column chromatography, such as solvent distillation are carried out using 60 silica of 230-400 mesh Kieselgel Elution.Ratio for TLC and the solvent of column chromatography is indicated with v/v.Pass through the UV light or use vanillic aldehyde or cerous molybdate of 254nm Dyeing visualizes compound.Unless otherwise stated, commercial materials use as it is.DCM is distilled out from calcium hydride And MeOH, THF and ether are distilled out from sodium/benzophenone.Tert-butanol is before use, in activationIt is dry on molecular sieve At least 24 hours.Anhydrous DMF is purchased from Sigma Aldrich.
Facility information
Unless otherwise stated, under the frequency of 300.2,400.2,500.2 and 600.2MHz, under 300K, Bruker is used Avance DPX 300, DPX 400, DPX 500 and DPX 600NMR spectrometer, record1H NMR spectra.1The displacement of H nmr chemical With hundred a ten thousandths (ppm) report, and refer to dissolvent residual signal: CDCl37.26, MeOD 3.31, δ acetone δ-d62.05 DMSO-δd62.50 δ and D2O 4.79。δ1H NMR data is reported as chemical shift (δH), relative integral, (s=is mono- for multiplicity Peak, d=doublet, t=triplet, q=quartet, two doublets of dd=, ddd=bimodal bimodal bimodal, dt=two A triplet, two triplets of td=, tri- triplets of tt=, two quartets of qd=), coupling constant (JHz) and to the greatest extent may be used Peak can be belonged to.In the presence of rotational isomer, when the ratio of rotational isomer is less than 1.5:1, it was recently reported that two kinds of rotations Turn isomers1H and13C NMR data.The case where 1.5 are greater than or equal to for ratio, only report main rotational isomeric Body.
Low resolution mass spectrometry is recorded on Finnigan LCQ Deca ion trap mass spectrometer (ESI).In Bruker Fu 7T High resolution mass spec is recorded on vertical leaf transformation ion cyclotron resonance mass spectrometer (FTICR).
Use the full-automatic melting point apparatus of Optimelt (Stanford Research Systems OptiMelt Automated Melting Point System) record fusing point.There is decaying total reflection (ATR) ability using 6.5 software of OPUS Infrared (IR) absorption spectrum is recorded on Bruker ALPHA spectrometer.It is measured using 341 polarimeter of Perkin Elmer Model Optical activity, and 10-1deg cm2g1Middle reportValue.-
Using with 2996 photodiode array detector of Waters or Waters 490E programmable wavelength detector 600 multi-solvent transportation system of Waters and Waters 500 pump, and use, operate under 254 and 280nm, use Sunfire Prep C18 OBD, 19x 50mm chromatographic column carries out preparative reversed-phase HPLC, and operation flow velocity is 7mL min1.Compound 0.1%TFA Or aqueous formic acid (solvent A) and 0.1%TFA or formic acid are in CH3Elution, uses the linear ladder of 0-50%B in CN (solvent B) Degree, it is more than 40 minutes that elution 40 minutes or 0-50%B, which is more than 45 minutes or 50-100%B,.
LC-MS is carried out on 2020 instrument of Shimadzu LC-MS, which is pumped by LC-M20A and be coupled to holotype The SPD-20A UV/Vis detector composition of 2020 mass spectrograph of Shimadzu (ESI) of formula operation.In Waters Sunfire5 μ It is separated on m, 2.1 × 150mm column (C18), with 0.2mL min-1Operated in flow rate.Use 0.1% aqueous formic acid (solvent A) and 0.1% formic acid is in CH3Mobile phase in CN (solvent B) is separated, linear gradient 0-50%B, and 30 minutes or 50- 100%B, 30 minutes.
Synthesis
The synthesis of uridine amine 18
The synthesis of uridine amine uses the Boojamra et al. modified1The route of previous publications carries out.
Scheme the synthesis of S1 uridine amine 18.Reagent and condition: a) TIPSOTf, iPr2NEt, DMF, room temperature, 2.5 hours, 82%；B) 10%Pd/C, H2(1atm), MeOH, room temperature, 30 minutes, 80%；C) (i) TFA:CH2Cl2(9:1v/v), room temperature, 15 Minute；(ii) isobutyl chlorocarbonate, iPr2NEt, THF, 0 DEG C to room temperature, 1.5 hours；(iii)NaBH4, H2O (dropwise addition), 0 DEG C, 1 Hour, 69%；D) toluene sulfochloride, pyridine, room temperature, 18 hours, 82%；e)NaN3, DMF, 75 DEG C, 4 hours, 82%；f)(i) TBAF (THF solution of 1M), THF, room temperature, 1 hour, (ii) Dowex 8-400, CaCO3, MeOH, room temperature, 1 hour, quantitatively； G) 1,3- dimercaptopropane, Et3N, MeOH, room temperature, 16 hours, 68%.
Tert-butyl (4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- ((triisopropyl first silicon Alkyl) oxygroup) -4,5- dihydrofuran -2- carboxylate (S2)
DMF (37mL) to ester S1 (3.64g, 12.3mmol) and n,N-diisopropylethylamine (2.1mL, 12.3mmol) is molten Triisopropylsilyl triflate (4.9mL, 17.6mmol) is added in liquid and is reacted.At room temperature, it stirs 2.5 hour.With saturation Na2CO3Aqueous solution (40mL) quenching reaction, and in Et2O (200mL) and H2It is distributed between O (40mL).It will Organic layer 0.2M HCl (50mL), H2O (5 × 50mL), salt water (50mL) washing, and through anhydrous Na2SO4It is dry.By organic layer Vacuum concentration, obtains thick residue, it is obtained the ester of white foam by column chromatography purifying (2:1v/v hexane: EtOAc) S2 (4.57g, 82%).
1H NMR (500MHz, CDCl3): δ 8.70 (s, 1H, NH), 7.03 (d, J=8.1Hz, 1H, H-6), 6.27 (d, J=3.2Hz, 1H, H-3 '), 5.96 (d, J=2.6Hz, 1H, H-1 '), 5.77 (d, J=8.1Hz, 1H, H-5), 5.23 (dd, J=3.3,2.6Hz, 1H, H-2 '), 1.54 (s, 9H, CO2 tBu), 1.12-1.07 (m, 3H, [CH (CH3)2]3Si), 1.06-1.02 (m, 18H, [CH (CH3)2]3Si)。13C NMR (126MHz, CDCl3): 162.5 (C of δ =O), 158.2,151.3,149.3,140.0,111.2,103.7,95.1,83.5,79.7,28.0,17.8,17.8,12.0. IR (ATR): 2944,2868,1725,1698cm-1。LRMS[M+H+]453.0。HRMS(ESI m/z)[M+Na+] C22H36N2O6SiNa calculated value, 475.2240；Measured value 475.2238.
Tert-butyl (2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- ((triisopropyl first Siloxy) tetrahydrofuran -2- carboxylate (S3)
10%Pd/C (86mg) is added into MeOH (21mL) solution of ester S2 (568mg, 1.25mmol), by alternately high Vacuum and nitrogen carry out 3 circulations, and reaction is made to deaerate.Under a hydrogen atmosphere by reaction mixture, it stirs 30 minutes.At this point, by anti- Object is answered to pass throughFiltering, and filter cake is thoroughly washed with MeOH (100mL).Solvent is removed in vacuum, obtains thick residue, Viscous, colorless oily ester S3 (480mg, 80%) is obtained by column chromatography purifying (1:1v/v hexane: EtOAc).
1H NMR (500MHz, CDCl3): δ 7.18 (d, J=8.1Hz, 1H, H-6), 5.72-5.68 (m, 2H, H-1 '+H-5), 4.78-4.74 (m, 2H, H-2 '+H-4 '), 2.57 (ddd, J=13.2,8.7, 6.2Hz, 1H, H-3 '), 2.24 (ddd, J=13.3,5.2,3.8Hz, 1H, H-3 '), 1.49 (s, 9H, CO2 tBu), 1.15-1.05 (m, 3H, [CH (CH3)2]3Si), 1.05-1.01 (m, 18H, [CH (CH3)2]3Si)。13C NMR (126MHz, CDCl3): δ 170.0 (C=O), 163.0 (C=O), 149.7 (C=O), 141.4,102.2,97.2,82.2,78.3,75.1,37.8,28.0, 17.8,17.8,11.9.IR (ATR): 2943,2867,1688cm-1。LRMS[M+H+]455.3。HRMS(ESI m/z)[M+Na+] C22H38N2O6SiNa calculated value, 477.2397；Measured value 477.2400.
1- ((2R, 3R, 5R) -5- (methylol) -3- ((triisopropylsilyl) oxygroup) tetrahydrofuran -2- base) pyrimidine -2,4 (1H, 3H)-diketone (S4)
Ester S3 (456mg, 1.0mmol) is dissolved in 9:1v/v TFA:CH2Cl2In (2.2mL), and at room temperature by reactant Stirring 15 minutes.Solvent is removed in vacuum, obtains residue, be then dissolved in THF (13mL) and is cooled to 0 DEG C, then Isobutyl chlorocarbonate (390 μ L, 3mmol) and n,N-diisopropylethylamine (260 μ L, 1.5mmol) is added.By reaction mixture plus Heat is stirred for 1.5 hours to room temperature, is subsequently cooled to 0 DEG C.It is added sodium borohydride (265mg, 7mmol), then at 20 minutes Interior dropwise addition water (2.3mL) is to dissolve sodium borohydride.Reaction mixture is stirred 1 hour at 0 DEG C, is then concentrated in vacuo.It will be residual Excess distributes between EtOAc (40mL) and water (10mL).Organic phase is separated, with 1M HCl (10mL), salt water (10mL) is washed, With anhydrous MgSO4It is dry.Solvent is removed in vacuum, obtains thick residue, by column chromatography (4:1v/v EtOAc: hexane → EtOAc white foam alcohol S4 (265mg, 69%)) is obtained.
1HNMR (400MHz, CDCl3): δ 8.94 (s, 1H), 7.25 (d, J= 8.1Hz, 1H, H-6), 5.72 (d, J=8.1Hz, 1H, H-5), 5.69 (d, J=2.1Hz, 1H, H-1 '), 4.73 (ddd, J= 5.7,3.5,2.1Hz, 1H, H-2 '), 4.61 (app.dq, J=9.0,4.7Hz, 1H, H-4 '), 3.80-3.72 (m, 2H, H- 5 '), 2.60 (s, 1H, OH), 2.27 (ddd, J=13.9,8.3,5.8Hz, 1H, H-3 '), 1.96 (ddd, J=13.6,5.0, 3.4Hz, 1H, H-3 '), 1.20-1.08 (m, 3H, [CH (CH3)2]3Si), 1.06 (d, J=7.0Hz, 18H, [CH (CH3)2]3Si)。13CNMR (101MHz, CDCl3): δ 163.2 (C=O), 150.0 (C=O), 140.2,102.1,95.3,82.4,76.1, 65.1,34.8,17.9,17.8,12.0.IR (ATR): 2943,2866,1686cm-1。LRMS[M+Na+]407.0。HRMS (ESIm/z)[M+Na+]C18H32N2O5SiNa calculated value, 407.1978；Measured value 407.1973.
((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- ((triisopropylsilyl) oxygen Base) tetrahydrofuran -2- base) methyl 4- toluenesulfonate (S5)
Alcohol S4 (1.53g, 3.97mmol) and p-toluenesulfonyl chloride (3.70g, 19.3mmol) are dissolved in pyridine (33mL) In, and reactant is stirred at room temperature 18 hours.At this point, reactant is concentrated in vacuo, residue is obtained, is dissolved in In EtOAc (200mL), with water (50mL), 1M HCl (50mL), salt water (50mL) washing and with anhydrous MgSO4It is dry.Vacuum is removed Solvent is removed, thick residue is obtained, colorless oil tosylate S5 is obtained by column chromatography purifying (1:1v/v hexane: EtOAc) (1.73g, 82%).
1HNMR (500MHz, CDCl3): δ 8.86 (s, 1H, NH), 7.80 (app.D, J=8.0Hz, 2H, Ar-H), 7.34 (d, J=8.0Hz, 2H, Ar-H), 7.13 (d, J=8.1Hz, 1H, H-6), 5.70 (d, J=8.1Hz, 1H, H-5), 5.54 (d, J=2.0Hz, 1H, H-1 '), and 4.75-4.67 (m, 1H, H-4 '), 4.65 (app.dt, J=5.1,2.0Hz, 1H, H-2 '), 4.21 (dd, J=10.4,7.0Hz, 1H, H-5 '), 4.12 (dd, J=10.4, 4.5Hz, 1H, H-5 '), 2.45 (s, 3H, CH3), 2.22 (ddd, J=13.7,8.3,5.4Hz, 1H, H-3 '), 1.88 (app.dt, J=13.7,3.6Hz, 1H, H-3 '), 1.15-1.02 (m, 3H, [CH (CH3))2]3Si), 1.00 (d, J=6.7Hz, 18H, [CH (CH3)2]3Si)。13CNMR (126MHz, CDCl3): δ 163.0,149.8,145.0,140.0,132.7,129.9, 128.0,102.1,95.7,79.0,75.8,71.5,35.2,21.7,17.9,17.8,11.9 .IR (ATR): 2944,2866, 1686cm-1。LRMS[M+Na+]561.0。HRMS(ESIm/z)[M+Na+]C25H38N2O7SSiNa calculated value, 561.2067；Measurement Value 561.2061.
1- ((2R, 3R, 5R) -5- (azido methyl) -3- ((triisopropylsilyl) oxygroup) tetrahydrofuran -2- base) is phonetic Pyridine -2,4 (1H, 3H)-diketone (S6)
Tosylate S5 (1.65g, 3.07mmol) is dissolved in DMF (34mL).Addition sodium azide (2.0g, 30.7mmol), reaction mixture is heated to 75 DEG C, is kept for 4 hours.Will reaction with EtOAc (200mL) dilute, with water (5 × 40mL), salt water (40mL) washs, and through anhydrous MgSO4It is dry.Solvent is removed in vacuum, obtains thick residue, it is passed through into column color It composes (1:1v/v EtOAc: hexane), obtains colorless oil azide S6 (1.09g, 87%).
1HNMR (400MHz, CDCl3): δ 9.01 (s, 1H, NH), 7.21 (d, J =8.1Hz, 1H, H-6), 5.72 (d, J=8.1Hz, 1H, H-5), 5.66 (d, J=2.1Hz, 1H, H-1 '), 4.75 (ddd, J= 5.7,3.6,2.1Hz, 1H, H-2 '), 4.62 (tt, J=7.4,4.7Hz, 1H, H-4 '), 3.60 (dd, J=12.7,7.1Hz, 1H, H-5 '), 3.39 (dd, J=12.7,4.6Hz, 1H, H-5 '), 2.28 (ddd, J=13.5,7.9,5.7Hz, 1H, H-3 '), 1.93 (ddd, J=13.5,4.9,3.5Hz, 1H, H-3 '), 1.19-1.08 (m, 3H, [CH (CH3)2]3Si), 1.06 (d, J= 6.7Hz, 18H, [CH (CH3)2]3Si)。13CNMR (101MHz, CDCl3): δ 163.2 (C=O), 149.9 (C=O), 140.4, 102.1,96.1,80.6,76.1,55.1,36.3,17.9,17.9,12.0.IR (ATR): 2944,2867,2102,1686cm-1。 LRMS[M+H+]410.0。HRMS(APCIm/z)[M+Na+]C18H31N5O4SiNa calculated value, 432.2043；Measured value 432.2038。
1- ((2R, 3R, 5R) -5- (azido methyl) -3- hydroxyl tetrahydrofuran -2- base) pyrimidine -2,4 (1H, 3H)-diketone (S7)
By the THF solution (280 μ L, 0.28mmol, 1.15eq.) of 1M TBAF be added drop-wise to azide S6 (0.10g, 0.24mmol, 1eq.) THF (1.8mL) solution in and at room temperature, stir 1 hour.Then by reaction mixture MeOH (2.8mL) dilution, and be stirred for 1 hour at room temperature with calcium carbonate (0.39g) and Dowex8-400 (1.2g).It then will be anti- It answers mixture to be filtered and concentrated in vacuo through Celite, obtains required colorless oil azide S7 (53.5mg, 88%), no It is used through being further purified.
[α]D 25=+25.8 ° (in CH2Cl2Middle c=0.21).1HNMR (300MHz, CDCl3): δ 10.53 (br, 1H, NH), 7.35 (d, J=7.5Hz, 1H, H-6), 5.81 (m, 1H, H-5), 5.72 (d, J=7.5Hz, 1H, H-1 '), 4.77-4.74 (m, 2H, H-2+H-4 '), 3.64 (m, 1H, H-5 '), 3.42 (m, 1H, H-5 '), 2.39 (m, 1H, H-3 '), 1.99 (m, 1H, H-3 '). IR (ATR): 3056,2101,1682,1461cm-1。LRMS[M+H+]253.1.These data and Boojamraet1Et al. report Data it is consistent.
1- ((2R, 3R, 5R) -5- (amino methyl) -3- hydroxyl tetrahydrofuran -2- base) pyrimidine -2,4 (1H, 3H)-diketone (18)
1,3- the third two is added in the solution in MeOH (1.0mL) to azide S7 (50.0mg, 198 μm of ol, 1eq.) Mercaptan (200 μ L, 2.0mmol, 10eq.) and Et3N (210 μ L, 1.5mmol, 7.6eq.).Reactant is stirred at room temperature 16 Hour, solvent is then removed in vacuum.Obtained residue is re-dissolved in H2In O (45mL), and use CH2Cl2(10×6mL) Washing.Water layer is lyophilized, required white fluffy solid amine 18 (46.4mg, 0.134mmol, 68%) is obtained, without into one Step is purified and is used.
1HNMR (500MHz, CD3OD): δ 7.56 (d, J=8.0Hz, 1H, H- 6), 5.75 (d, J=2.2Hz, 1H, H-1 '), and 5.69 (d, J=8.0Hz, 1H, H-5), 4.68-4.48 (m, 1H, H-4 '), 4.42 (ddd, J=6.1,3.6,2.2Hz, 1H, H-2 '), 2.91 (dd, J=13.6,3.7Hz, 1H, H-5 '), 2.83 (dd, J= 13.5,5.9Hz, 1H, H-5 '), 2.34 (ddd, J=14.1,8.1,6.2Hz, 1H, H-3 '), 1.81 (ddd, J=13.7,4.8, 3.6Hz, 1H, H-3 ').IR (ATR): 3375,1684,1629cm-1。LRMS[M+H+]228.0.These data with before by Boojamra et al.1The data of report are consistent.
The synthesis of the DABA segment of S8 and 20
Scheme the synthesis of the DABA segment of S2.S8 and 20.Reagent and condition: a) i.TFA:CH2Cl2(1:1v/v), room temperature, 30 Minute, ii.Fmoc-OSu, THF:H2O (1.2:1v/v), NaHCO3, two steps 70%；B) nano particle Zn, 2MHCl, MeOH, room Temperature, 16 hours, 78%；C) i.20 (1eq.), 10%Na2CO3, 0 DEG C, ii. allyl chlorocarbonate (1.1eq.), dioxanes, 0 DEG C To room temperature, 1.5 hours, 83%；D) i.2- chlorine trityl chloride resin, iPr2NEt, room temperature, 16h；Ii.20vol.% piperidines DMF solution, room temperature, 2 × 4 minutes.
(2S, 3S) -2- ((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino -3- ((benzyloxy) (methyl) amino) butyric acid (S10)
By sour S9 (according to previously by Boojamra et al.1The step of report, synthesizes) (4.42g, 13mmol) in TFA and CH2Cl2It synthesizes, is stirred at room temperature 30 minutes in the mixture of (1:1v/v, 30mL).Solvent is removed in vacuum, obtains residue, It is dissolved in THF (70mL).Saturation NaHCO is added3Aqueous solution (70mL) and solid NaHCO3(about 10g) is (mixed until reacting Object is closed in alkalinity), Fmoc- succinimide (4.62g, 14mmol) then is added.Reaction is stirred at room temperature 16 hours.This When, H is added2O (100mL) simultaneously washs reaction mixture with ether (3 × 50mL).Water layer is acidified to pH2 simultaneously with 1M HCl It is extracted with EtOAc (3 × 250mL).The anhydrous MgSO of combined organic layer4It is dry, solvent is removed in vacuum, obtains thick residue, It is obtained into colorless oil Fmoc- protection by column chromatography purifying (3:1v/v hexane: EtOAc → 1:1v/v hexane: EtOAc) Sour S10 (4.19g, 70%).
1HNMR(500MHz,CDCl3):δ7.96-7.69(m,2H,Ar-H), 7.59 (dd, J=7.7,3.4Hz, 2H, Ar-H), 7.41-7.27 (m, 9H, Ar-H), 5.60 (d, J=8.3Hz, 1H, NH), 4.75(s,2H,CH2Ph),4.43(m,3H,α-CH+Fmoc-CH2), 4.23 (app.t, J=7.2Hz, 1H, Fmoc-CH), 3.30-3.05(m,1H,β-CH),2.68(s,3H,NCH3), 1.22 (d, J=6.9Hz, 3H, γ-CH3).13CNMR(126MHz, CDCl3):δ173.5,156.8,143.8,141.5,136.0,129.0,128.8,128.6,127.3,127.2,125.3, 120.1,74.7,67.4,64.2,55.3,47.3,41.2,10.1.IR(ATR):2955,1720cm-1.LRMS[M+H+] 461.4.HRMS(ESIm/z)[M+Na+]C27H28N2O5Na calculated value, 483.1896；Measured value, 483.1890.
(2S, 3S) -2- ((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino) -3- (methylamino) butyric acid (S8)
Nano particle zinc (2.15g, 33mmol) is added to the MeOH for the sour S10 (3.04g, 6.6mmol) being vigorously stirred In (34mL) solution.2M HCl (34mL) solution is added dropwise, and reactant is stirred at room temperature 2 hours.Then it is added another A part of nano particle zinc (0.31g, 4.8mmol) and 2M HCl (11mL), and reactant is stirred at room temperature 16 hours.It will Reactant is concentrated in vacuo to the half of its volume, then uses H2O (400mL) dilution, and use CH2Cl2(80mL) washing.By water layer It is extracted with EtOAc (5 × 150mL) and EtOAc/MeOH mixture (9:1v/vEtOAc:MeOH, 2 × 150mL).By having for merging The anhydrous MgSO of machine layer4It is dry, solvent is removed in vacuum, obtains white solid acid S8 (1.82g, 78%), without further pure Change and uses.
133-135 DEG C of fusing point (decomposition).1HNMR(500MHz,CD3OD):δ 7.86-7.76 (m, 2H, Ar-H), 7.70 (app.t, J=7.4Hz, 2H, Ar-H), 7.40 (dd, J=8.1,6.7Hz, 2H, Ar- ), H 7.32 (ddd, J=7.7,6.8,1.6Hz, 2H, Ar-H), 4.72-4.67 (m, 1H, DABA- α-CH), 4.55 (dd, J= 10.5,6.7Hz,1H,Fmoc-CH2),4.41-4.32(m,1H,Fmoc-CH2), 4.27 (app.t, J=6.7Hz, 1H, Fmoc- CH),3.77-3.70(m,1H,DABA-β-CH),2.75(s,3H,NCH3), 1.26 (d, J=6.5Hz, 3H, DABA- γ-CH3) .13CNMR(100MHz,CDCl3): δ 174.2 (C=O), 156.7 (C=O), 144.0,141.4,136.2,129.0,128.6, 128.4,127.8,127.2,125.2,120.0,74.5,67.3,64.1,55.7,47.2,41.4,14.3.IR(ATR): 3335,3049,1704cm-1.LRMS[M+H+]355.1.HRMS(ESIm/z)[M+Na+]C27H28N2O5Na calculated value, 483.1896；Measured value, 483.1890.
(2S, 3S) -2- (((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino) -3- (((allyloxy) carbonyl) amino) Butyric acid (20)
Sour S8 (0.60g, 1.7mmol, 1eq.) is dissolved in 10% aqueous sodium carbonate (14.3mL) and is cooled to 0 ℃.Isosorbide-5-Nitrae-dioxanes (6.2mL) solution of allyl chlorocarbonate (196uL, 1.9mmol, 1.1eq.) is added drop-wise to above-mentioned reaction In mixture, and reaction mixture is heated to room temperature and is stirred 1.5 hours.Then use H2O (120mL) diluting reaction, uses second Ether (2 × 40mL) washs and is acidified to pH 2 with 1M HCl.Then EtOAc (3 × 200mL) aqueous layer extracted, organic phase warp are used MgSO4It is dry, merge and be concentrated in vacuo yellowly grease, contains required white foam acid 20 (0.61g, 83%).1HNMR(400MHz,CDCl3): δ 7.75 (d, J=7.5Hz, 2H, Ar-H), 7.61-7.55 (m, 2H, Ar-H), 7.39 (app.t, J=7.5Hz, 2H, Ar-H), 7.30 (app.t, J=7.5Hz, 2H, Ar-H), 5.97-5.81 (m, 2H, NH+Alloc- β-CH), 5.33-5.14(m,2H,Alloc-γ-CH2),4.73-4.32(m,6H,Alloc-α-CH2+Fmoc-CH2+DABA-α-CH+ DABA- β-CH), 4.20 (t, J=6.8Hz, 1H, Fmoc-CH), 2.85 (s, 3H, NCH3), 1.25 (d, J=6.7Hz, 3H, DABA-γ-CH3).13CNMR(101MHz,CDCl3): δ 173.0 (C=O), 156.1 (C=O), 143.8,143.6,141.3, 132.5,127.7,127.1,125.0,120.0,117.6,67.2,66.6,57.1,53.4,47.2,29.5,14.1.IR (ATR):3312,2984,1688,1683,1528cm-1.LRMS[M+H+]439.3.HRMS(ESIm/z)[M+Na+] C24H26N2O6Na calculated value, 461.1688；Measured value, 461.1683.
(2S, 3S) -3- ((allyloxy) carbonyl) amino) -2-amino-butyric acid is on 2- chlorine trityl chloride resin (S11)
16 hours at room temperature, amino acid 20 (515mg, 1.1mmol) is loaded into CH2Cl22- chlorine three in (6.5mL) On benzyl chlorine resin (745mg, 1.05mmol).According to general step 2, with n,N-diisopropylethylamine (550 μ L, It 3.1mmol) is deprotected with Fmoc-, obtains S11 (at λ=301nm spectral measurement fulvene-piperidine adduct of resin-bonded The load capacity 65% of measurement).
The synthesis of tryptophan 4- nitrophenylcarbamate S12
Scheme the synthesis of S3. tryptophan 4- nitrophenylcarbamate S12.Reagent and condition: a) 20vol.% in MeCN Piperidines, room temperature, 30 minutes, 75%；B) 4- chloroformate nitrophenyl ester, CH2Cl2, room temperature, 18 hours, 80%.
Tert-butyl (S) -3- (3- (tertbutyloxycarbonyl) -2- (((4-nitrophenoxy) carbonyl) amino) -3- oxopropyl) - 1H- indoles -1- carboxylate (S12)
By acetonitrile (20mL) the solution processing of tert-butyl ester S13 (2.76g, the 4.74mmol) piperidines of 20vol.%, and At room temperature by reactant, it stirs 20 minutes.Solvent is removed in vacuum, obtains thick residue, by its by column chromatography purifying (3: 1v/v hexane: EtOAc, 0.1vol.%Et2NH), the tryptophan yellow oily S14 (1.28g, 75%) being deprotected.To The CH of S14 (1.28g, 3.56mmol) and n,N-diisopropylethylamine (620 μ L, 3.56mmol)2Cl2It is added in (26mL) solution Reactant is stirred at room temperature 19 hours rubigan chloro-formate (860mg, 4.27mmol).Solvent is removed in vacuum, obtains Thick residue obtains white foam amino by column chromatography purifying (7:1v/v hexane: EtOAc → 5:1v/v hexane: EtOAc) Formic acid esters S12 (1.49g, 80%).
1HNMR(400MHz,CDCl3): δ 8.23 (d, J=9.1Hz, 2H, H-3 + H-5), 7.57 (app.dt, J=7.9,1.0Hz, 1H, Ar-H), 7.46 (s, 1H, Ar-H), 7.34 (ddd, J=8.4,7.2, 1.3Hz, 1H, Ar-H), 7.29-7.21 (m, 3H, Ar-H), 5.74 (d, J=7.8Hz, 1H, NH), 4.70-4.58 (m, 1H, α- ), CH 3.35 (dd, J=14.8,5.8Hz, 1H, β-CH2), 3.25 (dd, J=14.9,5.5Hz, 1H, β-CH2),1.66(s,9H, 3×CH3),1.46(s,9H,3×CH3).13CNMR(101MHz,CDCl3): δ 170.0 (C=O), 155.7 (C=O), 152.5 (C =O), 144.9,130.7,125.5,125.1,124.7,124.2,122.7,122.0,121.6,11 8.9,115.4,114.8, 83.8,83.1,54.9,28.2,28.0,27.5.IR(ATR):2988,2973,1724cm-1.LRMS[M(-NO2Ph+Me)+Na+]441.0.HRMS(ESIm/z)[M(-NO2Ph+Me)+Na]+Calculated value C22H30N2O6Na441.2002；Measured value, 441.1995。
Scheme the synthesis 87 of S4Fmoc- uridine amine.
1- ((3aR, 4R, 6R, 6aR) -6- (methylol) -2- methoxyl group tetrahydrofuran [3,4d] [1,3] dioxole 4- yl) pyrimidine -2,4 (1H, 3H)-diketone (84)
P-methyl benzenesulfonic acid is added into trimethyl orthoformate (13mL) solution of uridine (5.0g, 21mmol, 26eq.) (0.14g, 1.6mmol, 1eq.), and be stirred at room temperature 16 hours.Then the reaction is cooled to 0 DEG C, 28%w/v is then added The MeOH solution (0.15mL) of NaOMe, is subsequently added into toluene (5mL), and at 0 DEG C, is stirred for 1 hour.Then reaction is mixed It closes object to filter and washed with toluene, obtains the pure white precipitate 84 (5.5g, 19mmol, 94%) of diastereo-isomerism.
1HNMR(400MHz,(CD3)2SO) δ 11.38 (s, 1H, NH), 7.77 (d, J=8.0Hz, 1H, H-6), 6.09 (s, 1H, CH), 5.80 (d, J=2.7Hz, 1H, H-1 '), 5.63 (d, J=8.0Hz, 1H, H-5), 5.09 (t, J=5.4Hz, 1H, OH), 5.00 (dd, J=6.5,2.6Hz, 1H, H-2 '), 4.85 (dd, J=6.4,3.8Hz, 1H, H-3 '), 4.06 (dt, J= 4.5Hz, 1H, H-4 '), 3.59 (dd, J=11.9,5.3Hz, 2H, CH2),3.21(s,3H,CH3)；13CNMR(100MHz, (CD3)2SO)δ163.7,150.8,142.6,117.3,102.2,91.5,86.2,83.2,80.5,61.6,50.7；LRMS[M+ H]+287.0.These data and Yasuda et al.[1]The data of report are consistent.
1- ((2R, 3R) -5- ((benzhydryl amino) methyl) -3- hydroxyl -2,3-dihydrofuran -2- base) pyrimidine -2,4 (1H, 3H)-diketone (85)
DMSO (0.43mL, 6mmol, 6eq.) is added into DMF (6.3mL) solution of 84 (0.29g, 1mmol, 1eq.), EDCHCl (0.58g, 3mmol, 3eq.) is subsequently added into DMF (1.9mL), premix TFA (40 μ L, 2mmol, 2eq.) and The premix of pyridine (80 μ L, 1mmol, 1eq.).By reaction stirring 2 hours, Et is then added3N (0.56mL, 4mmol, 4eq.) and it is stirred for 30 minutes.Then oxalic acid (0.25g, 0.5mmol, 0.5eq.) is added, the remnants that then vacuum will obtain Object is re-dissolved in CH2Cl2In 10%v/vMeOH in, and filtered by silica plug.Solvent is removed in vacuum, obtains yellow Grease is then re-dissolved in MeOH (2.8mL).Thereto be added benzhydryl amine (0.19mL, 1.1mmol, 1.1eq.), NaCNBH3(94mg, 1.5mmol, 1.5eq.) is then added AcOH (40 μ L, 0.7mmol, 0.7eq.) and stirs 15 hours.Solvent is removed in vacuum, obtained residue is re-dissolved in CH2Cl2In, with saturation NaHCO3Aqueous solution, salt washing It washs, dry (MgSO4).Organic phase is concentrated in vacuo, and by column chromatography eluting residue, with 1:4v/v hexane: EtOAc is washed It is de-, it is then eluted with pure EtOAc, obtains yellow colored foam 85 (0.12g, 0.29mmol, 29%, 2 steps).
1HNMR(400MHz,CDCl3)δ7.42-7.39(m,4H,Ar-H),7.33-7.30(m,4H,Ar-H),7.25- 7.21 (m, 2H, Ar-H), 7.19 (d, J=8.1Hz, 1H, H-6), 6.27 (d, J=1.45Hz, 1H, H-1 ') 5.64 (d, J= 8.1Hz, 1H, H-5), 5.21 (d, J=2.3Hz, 1H, H-3 '), 4.92 (s, 2H, CH+H-2 '), 3.40 (s, 2H, CH2)；13CNMR(100MHz,CDCl3)δ163.4,161.2,150.6,143.0,138.9,128.7,128.6,127.4,127.3, 127.2,127.1,103.0,100.7,93.3,79.7,66.4,44.1；HRMS calculated value C22H21N3O4: MNa+, 414.14243.Measured value: MNa+,414.14247。
1- ((2R, 3R) -5- ((benzhydryl amino) methyl) -3- ((triisopropylsilyl) oxygroup) -2,3- dihydro furan Mutter -2- base) pyrimidine -2,4 (1H, 3H)-diketone (86)
To 85 (0.12g, 0.29mmol, 1eq.) and iPr2The DMF (0.8mL) of EtN (56 μ L, 0.32mmol, 1.1eq.) TIPS-OTf (0.12mL, 0.44mmol, 1.5eq.) is added dropwise in solution, and stirs 1 hour.Reaction mixture is concentrated in vacuo, Obtained residue is re-dissolved in CH2Cl2In.By organic phase saturation NaHCO3Aqueous solution, H2Then O is washed with brine, Na2SO4It is dry, and be concentrated in vacuo.Gained residue is purified by column chromatography, and with 2:1v/v hexane: EtOAc is eluted, and obtains white Foam 86 (0.13g, 0.24mmol, 84%).
1HNMR(400MHz,CDCl3)δ7.43-7.41(m,4H,Ar-H),7.35-7.31(m,4H,Ar-H),7.27- 7.23 (m, 2H, Ar-H), 7.14 (d, J=8.2Hz, 1H, H-6), 6.35 (d, J=2.1Hz, 1H, H-1 '), 5.72 (d, J= 8.1Hz, 1H, H-5), 5.15 (d, J=2.3Hz, 1H, H-3 '), 5.05 (t, J=2.2Hz, 1H, H-2 '), 4.93 (s, 1H, CH),3.41(s,2H,CH2),1.10-1.08(m,21H,[CH(CH3)2]3)；13CNMR(100MHz,CDCl3)δ162.9, 161.0,149.7,143.2,143.1,139.5,128.6,127.3,103.4,101.4,93.3,80.4,66.1,44.2, 17.9,12.1；HRMS calculated value C31H42N3O4Si:MNa+,570.27585.Measured value: MNa+,570.27586。
(9H- fluorenes -9- base) methyl (((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- Hydroxyl tetrahydrofuran -2- base) methyl) carbamate (87)
Into MeOH (12mL) solution of 86 (0.13g, 0.24mmol, 1eq.) be added 10%Pd/C (51mg, 0.05mmol, 0.2eq.) and AcOH (69 μ L, 1.2mmol, 5eq.).Solvent is deaerated 10 minutes, then flask is evacuated and is used in combination N2(g)Backfill is three times.Then make reaction in H2(g)It is stirred 3.5 hours under atmosphere.After the reaction was completed by LCMS measurement, H is removed2(g) And reaction mixture is passed throughIt is filtered and concentrated in vacuo.Gained residue is re-dissolved in the 95%EtOH of 4M HCl It in (6mL) solution and stirs 3 hours, is then concentrated in vacuo, gained residue is then re-dissolved in H2In O, EtOAc is used (x3) it washs and is lyophilized.Residue is re-dissolved in 1:1v/v THF: saturation NaHCO3(aq)In (1.3mL).It is added into the solution Fmoc-OSu (48mg, 0.14mmol, 1.1eq.) is simultaneously stirred 16 hours.Then reaction mixture is diluted with EtOAc, uses H2O, Salt water washing, dry (MgSO4) and be concentrated in vacuo.Then by column chromatography eluting obtained thick residue, CH is used2Cl2It washes It is de-, then with the CH containing 10%MeOH2Cl2Elution, obtains 87 (41mg, 0.090mmol, 3 steps, 47%) of white foam.
1H NMR(400MHz,CDCl3)δ7.76-7.74(m,2H,Ar-H),7.60-7.58(m,2H,Ar-H),7.40- 7.36(m,2H,Ar-H),7.31-7.27(m,3H,2x Ar-H+1x H-6),5.77(m,2H,1x NH+1x H-1’),5.69 (d, J=7.9Hz, 1H, H-5), 5.06 (br s, 1H, OH), 4.53-4.50 (m, 2H, 1x H-2 '+1x H-4 '), 4.42- 4.37(m,2H,Fmoc-CH2), 4.21 (t, J=6.8Hz, 1H, Fmoc-CH), 3.52-3.39 (m, 2H, H-5 '), 2.23- 2.20(m,1H,H-3'),1.95-1.91(m,1H,H-3')；13C NMR(100MHz,CDCl3)δ163.9,156.7,151.3, 143.9,143.8,141.3,139.1,127.7,127.1,125.1,120.0,102.3,94.2,80.9,66.8,50.7, 47.2,45.4,33.7,29.7；LRMS[M+H]+450.0.These data and Tran[2]The data of report are consistent.
Scheme the synthesis of S5 connexon 88.
The synthesis of connexon 88 is synthesized by the method in the disclosure of Torres-Garcia etc.[3]
4- ((3,4- dihydro -2H- pyrans -2- base) methoxyl group) benzoic acid (88)
In THF (22mL), by 4-HBA methyl esters (2.0g, 13mmol, 1eq.) and PPh3(3.8g, 14mmol, Solution 1.2eq.) stirs 30 minutes.Then it at 0 DEG C, is added DIAD (2.8mL, 14mmol, 1.2eq.).Reaction is heated To room temperature and stir 16 hours.Solvent is removed in vacuum, is re-dissolved in 2:1v/v H2In O:MeOH.Addition LiOH (1.4g, 59mmol, 4.5eq.), mixture is flowed back 16 hours at 120 DEG C.Reaction mixture is cooled to room temperature, CH is then used2Cl2(x3) it washes It washs.Water phase is acidified to pH 2 using 1M HCl (aqueous solution), filtering gained white depositions are simultaneously dried in vacuo, and obtain white powder End 88 (3.7g, 16mmol, quantitative).
1H NMR(400MHz,(CD3)2SO)δ12.61(s,1H,COOH),7.91-7.87(m,2H,Ar-H),7.05- 7.01 (m, 2H, Ar-H), 6.40 (m, 1H, OCH=CH), 4.72-4.69 (m, 1H, OCH=CH), 4.13-4.12 (m, 3H, 1x CH+2x OCH2),2.10-2.03(m,1H,H-3),1.98-1.95(m,2H,1x H-3+1x H-4),1.74-1.64(m,1H, H-4)；13C NMR(100MHz,(CD3)2SO)δ167.0,162.0,143.2,131.4,123.2,114.3,100.5,72.8, 69.9,23.6,18.7.LRMS[M+H]+235.0.These data and TorresGarcia et al.[3]The data of report are consistent.
Scheme the synthesis in solid state of S6 uridine peptide antibiotics (sansanmycin) analog.
General step 1: the synthesis of different peptide
HATU (1eq.) is added into DMF (4-8mL/mmol) solution of boc-protected amino acid (1eq.), and in room Under temperature, stirs 10 minutes first, solution is then cooled to 0 DEG C.It is added S8 (1eq.), NMM or N, N- diisopropyl is then added Base ethamine (3eq.), is heated to room temperature for reaction mixture and stirs 3-6h.After the completion, by reaction mixture EtOAc (60- It 100mL) dilutes, and successively uses 0.2MHCl (15-20mL), water (5 × 15mL) and salt water (15mL) washing.Solvent is removed in vacuum, Thick residue is obtained, it is purified by column chromatography or reversed-phase HPLC, obtains different peptide S15-S16.
(2S, 3S) -2- ((((9H fluorenes -9- base) methoxyl group) carbonyl) amino) -3- ((S) -2- ((tertbutyloxycarbonyl) ammonia Base) -3- (3- ((tert-butyl dimetylsilyl) oxygroup) phenyl)-N- methyl propanamide base) butyric acid (S15)
According to general step 1, in the DMF (8.9mL) of HATU (433mg, 1.14mmol) and NMM (376 μ L, 3.42mmol) In the presence of solution, Boc-L-m-Tyr (OtBu)-OH (384mg, 1.14mmol) and S8 (406mg, 1.14mmol) react 6 hours, After column chromatography (1:1v/vEtOAc: hexane → EtOAc) separation, obtain the different peptide S15 of white, amorphous solid (384mg, 50%).
1HNMR(400MHz,CDCl3):δ7.81-7.68(m,2H,Ar-H), 7.64-7.53 (m, 2H, Ar-H), 7.43-7.34 (m, 2H, Ar-H), 7.33-7.27 (m, 2H, Ar-H), 7.08 (d, J= 7.8Hz, 2H, Ar-H, H-2+H-6), 6.94-6.83 (m, 2H, Ar-H, H-3+H-5), 5.91 (s, 1H, NH), 5.58 (d, J= 8.6Hz,1H,NH),4.92-4.83(m,1H,Tyr2-α-CH),4.80-4.67(m,1H,DABA1-β-CH),4.40-4.29 (m,3H,Fmoc-CH2+ DABA1- α-CH), 4.20 (app.t, J=6.7Hz, 1H, Fmoc-CH), 3.05-2.87 (m, 4H, NCH3 +Tyr2-β-CH2),2.81-2.68(m,1H,Tyr2-β-CH2),1.56-1.00(m,21H,Boc+tBu+DABA1-γ-CH3) .IR(ATR):3335,3045,1704cm-1.LRMS[M+H+]673.3.HRMS(ESIm/z)[M+Na+]C38H47N3O8Na is calculated Value, 696.3261；Measured value, 696.3266.
(2S, 3S) -2- ((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino) -3- ((S-2- ((tertbutyloxycarbonyl) ammonia Base)-N- methyl -3- phenylpropionyl amido) butyric acid (S16)
According to general step 1, in the DMF (1.1mL) of HATU (106mg, 0.28mmol) and NMM (9.2 μ L, 0.84mmol) In the presence of solution, react Boc-L-Phe-OH (70mg, 0.26mmol) and S8 (100mg, 0.28mmol) 4 hours, column chromatography (95:5v/v CH2Cl2: MeOH) after, obtain the different peptide S16 of white foam (73mg, 46%).
1HNMR(300MHz,CDCl3, major rotomer) and δ 7.82 (d, J=7.5Hz, 2H, 2xAr-H), 7.69 (m, 2H, 2xAr-H), 7.39-7.34 (m, 2H, 2xAr-H), 7.30-6.96 (m, 7H, 4xAr-H+H-2+H-3+H-5), 5.93 (d, J=8.4Hz, 1H, N-H), 5.78 (d, J=8.2Hz, 1H, N-H), 5.03 (m,1H,Phe2-α-CH),4.68(m,1H,DABA1-β-CH),4.53(m,1H,DABA1-α-CH),4.40-4.27(m,3H, Fmoc-CH2+Fmoc-CH),3.00-2.77(m,4H,NCH3+Phe2-β-CH2),2.74(m,1H,Phe2-β-CH2),1.34- 1.05(m,12H,Boc+DABA1-γ-CH3).IR(ATR):3316,2979,2919,1712,1640cm-1.LRMS[M+H+] 602.4.HRMS(ESIm/z)[M+Na+]C34H39N3O7Na calculated value, 624.2685；Measured value, 624.2683.
The synthesis of depsipeptide S24-S43, S45-S52
General step 2: amino acid is loaded on 2- chlorine trityl chloride resin
Condition A:
Make the 2- chlorine trityl chloride resin with 1%DVB (1.22-1.42mmol/g, 50-730 μm of ol, 1eq.) (100-200 mesh) is in anhydrous CH2Cl2Swelling 30 minutes in (3-8mL).By different peptide S15-S16 (100-320 μm of ol, 2eq.) or ammonia Base acid 20 (1.1-2.5mmol, 1.2eq.) is dissolved in anhydrous CH2Cl2(it is 2-2.6mL/100 μm of ol for different peptide S15-S16, for Amino acid 20 is 0.6mL/100 μm of ol).Addition n,N-diisopropylethylamine (it is 200-2920 μm of ol for different peptide S15-S16, 2-8eq. is 2.4-3eq. for amino acid 20), and resin is vibrated at room temperature 16 hours.DMF (5 × .5mL) then is used, CH2Cl2(5 × 5mL) and DMF (5 × 5mL) wash resin.With 17:2:1v/v/v CH2Cl2: methanol: at iPr2NEt (3-6mL) Reason 40 minutes, covers resin.Then DMF (5 × 5mL) is used, CH2Cl2(5 × 5mL) and DMF (5 × 5mL) wash resin.
Condition B:
DMF (x3) solution of Rink amide resin 20%v/v piperidines is handled, vibrates 3 minutes, then uses DMF (x5), CH2Cl2(x5) it is washed with DMF (x5).By in λ=301nm (ε=7800cm-1) under fulvene piperidine adduct spectrum survey Measure the load capacity to determine resin.By connexon 88 (0.062-0.21mmol, 4eq.), PyBOP (0.062-0.21mmol, 4eq.) it is added in resin and vibrates 1 hour with DMF (0.10M) solution of NMM (0.062-0.21mmol, 4eq.).Then use DMF (x5), CH2Cl2(x5) and DMF (x5) washs resin, then with the pyridine solution processing of 10%v/v acetic anhydride 3 minutes, together When shake, then use DMF (x5) and CH2Cl2(x10) it washs.Then by resin at 80 DEG C, at 1,2- dichloroethanes (0.1M) In flow back 16 hours together with 87 (0.038-0.13mmol, 2.5eq.) and PPTS (0.012-0.058mmol, 1.1eq.).With CH2Cl2(x5) and DMF (x5) washs resin, is then handled as described above with the DMF solution of 20% piperidines, measures its load effect Rate.
General step 3:Fmoc- strategy Solid phase peptide synthesis
Fmoc deprotection: 10vol% piperidines/DMF (5mL) solution is added into resin and vibrates 4 minutes (× 2).Then With DMF (5 × 3mL), CH2Cl2(5 × 3mL) and DMF (5 × 3mL) wash resin.Added by the fulvene piperidines at λ=301nm The spectral measurement of object is closed to determine the efficiency of previous amino acid couplings.
Amino acid couplings:
Condition A: by the amino acid (200-640 μm of ol, 4eq.) of protection, PyBOP (200-640 μm of ol, 4eq.) and NMM DMF (0.1M) solution of (0.4-1.28mmol, 8eq.) is added in resin (1eq.), and at room temperature, is vibrated 1 hour.So DMF (5 × 5mL) is used afterwards, CH2Cl2(5 × 5mL) and DMF (5 × 5mL) wash resin.
Condition B: by fmoc-protected amino acid (72-96 μm of ol, 1.2eq.), HOAt (72-96 μm of ol, 1.2eq.) and DMF (0.1M) solution of DIC (72-96 μm of ol, 1.2eq.) is added in resin (1eq.), and is vibrated 16 hours at room temperature. Then DMF (5 × 3mL) is used, CH2Cl2(5 × 3mL) and DMF (5 × 3mL) wash resin.
Capping: 10vol% acetic anhydride/pyridine (5mL) solution is added into resin, and vibrates 3 minutes.Then use DMF (5 × 5mL), CH2Cl2(5 × 5mL) and DMF (5 × 5mL) wash resin.
General step 4: solid phase urea synthesis
By carbamate S12 (90-320 μm of ol, 2eq.) and n,N-diisopropylethylamine (180-640 μm of ol, 4eq.) DMF (17 μ L/ μm ol) solution be added in resin (45-160 μm of ol, 1eq.).Resin is vibrated at room temperature 6 hours.With DMF (5 × 5mL) is used afterwards, CH2Cl2(5 × 5mL) and DMF (5 × 5mL) wash resin.
General step 5: the solid phase deprotection of allyl carbamate
By the CH of tetrakis triphenylphosphine palladium (0) (20 μm of ol, 0.2eq.) and phenyl silane (2mmol, 20eq.)2Cl2 (80mM) solution is added in resin (1eq.), and is shaken 15 minutes at room temperature.Solvent is then removed from resin, is used in combination CH2Cl2(5 × 5mL), DMF (5 × 5mL) and CH2Cl2(5 × 5mL) washs resin.Then reprocessing is primary.
General step 6: solid phase is coupled to N- methylated amino-acid residue
Amino acid couplings: boc-protected amino acid (80-580 μm of ol, 2eq.), HATU (80-580 μm of ol, 2eq.) and DMF (0.1M) solution of DIPEA (120-870 μm of ol, 2-3eq.) is added in resin (1eq.), and vibrates 2 hours at room temperature. Then DMF (5 × 5mL) is used, CH2Cl2(5 × 5mL) and DMF (5 × 5mL) wash resin.
Capping: pyridine (5mL) solution of 10vol% acetic anhydride is added into resin and vibrates 3 minutes.Then DMF (5 is used × 3mL), CH2Cl2(5 × 3mL) and DMF (5 × 3mL) wash resin.
General step 7: it cracks and is post-processed from 2- chlorine trityl chloride resin
By the CH of 30vol% hexafluoroisopropanol (HFIP)2Cl2(5-10mL) solution is added in resin, and is shaken at room temperature It swings 30 minutes.Then use CH2Cl2(6 × 10mL) washs resin, and merges lysate and cleaning solution, is concentrated in vacuo.Gained is residual Excess is dissolved in 9:1v/v MeCN:H2In O, and purified by reversed-phase HPLC.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- ((S) -2- ((uncle Butoxy carbonyl) amino)-N- methyl -3- phenylpropionyl amido) ethyl) tri- oxygen of -5- isobutyl group -12,12- dimethyl -4-, 7,10- Three azepine tridecane -1- of generation -11- oxa- -3,6,8- is sour (S24)
According to general step 2, it is deprotected using n,N-diisopropylethylamine (61 μ L, 350 μm of ol) and Fmoc- by different peptide S16 (105mg, 175 μm of ol), which is loaded to, is dissolved in CH2Cl22- chlorine trityl chloride resin (62mg, 88 μm of ol) in (2mL) On.Then by Fmoc-L-Leu-OH (124mg, 350 μm of ol) and PyBOP (182mg, 350 μm of ol) and NMM (77 μ L, 700 μ Mol) (general step 3) is coupled in DMF (1mL).Then according to general step 4 and carbamate S12 (92mg, 175 μ Mol it) is coupled.Then from resin crack (general step 7), by reversed-phase HPLC (50-100%MeCN, 40 minutes；50% MeCN, 10 minutes) purified peptide, obtain fluffy white solid depsipeptide (depsipeptide) S24 (12.4mg, 39%).
IR(ATR):3340,2976,2931,1730,1635cm-1.1HNMR(400MHz,CDCl3):δ8.12(1H,d,J =8.0Hz, Ar-H), 7.83 (1H, d, J=8.0Hz, Ar-H), 7.69 (1H, d, J=4.4Hz, Ar-H), 7.52 (1H, s, Ar- H),7.34-7.15(7H,m,7xAr-H),6.24-6.14(1H,m,N-H),6.07-5.87(1H,m,N-H),5.05(1H,m, DABA1-β-CH),4.80-4.66(3H,m,DABA1-α-CH+Phe2-α-CH+Trp4-α-CH),4.43-4.38(1H,m, Leu3-α-CH),3.20-3.06(5H,m,Trp4-β-CH2+NCH3),2.82–2.76(2H,m,Phe2-β-CH2),1.77- 1.48(12H,m,3xCH3+Leu3-β-CH2+Leu3-γ-CH),1.40-1.28(18H,m,6xCH3),1.23-1.18(3H,m, DABA1-γ-CH3),0.93–0.89(6H,m,2xLeu3-δ-CH3).LRMS[M+H+]879.5.HRMS(ESIm/z)[M+Na+] C46H66N6O11Calculated value 901.4687, measured value 901.4687.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -5- isobutyl group -12,12- diformazan Three azepine tridecanoic acid (S25) of base -2- ((S) -1- (methylamino) ethyl) -4,7,10- trioxy- -11- oxa- -3,6,8-
Using standard Fmoc-SPPS step, the DMF of PyBOP (1.64g, 3.16mmol) and NMM (650 μ L, 6.3mmol) (7.9mL) solution, Fmoc-L-Leu-OH (1.12g, 3.16mmol) and S11 (0.79mmol) coupling of resin-bonded is (general Step 3, condition A).Fmoc deprotection, then according to general step 4, using n,N-diisopropylethylamine (550 μ L, 3.16mmol) it is coupled carbamate S12 (840mg, 1.58mmol).Four (triphenylphosphines) are then used according to general step 5 Palladium (0) (2 × 194mg, 2 × 158 μm of ol) and phenyl silane (2 × 1.9mL, 2 × 15.8mmol) remove allyl amino formic acid Ester obtains the S25 with resin-bonded.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- ((S) -2- ((uncle Butoxy carbonyl) amino)-N- methyl-3- (pyridin-3-yl) propionamido) ethyl) dimethyl-4,7-5- isobutyl group-12,12-, Three azepine tridecanoic acid (S26) of 10- trioxy- -11- oxa- -3,6,8-
According to general step 6, in DMF (0.6mL), HATU (45mg, 120 μm of ol) and n,N-diisopropylethylamine are used The S25 (60 μm of ol) of resin-bonded and Boc-3-Pal-OH (32mg, 120 μm of ol) are coupled by (33 μ L, 180 μm of ol).? From resin crack after (general step 7), by reversed-phase HPLC (20-100%MeCN, 40 minutes；20%MeCN, 10 minutes) it is pure Change peptide, obtains fluffy white solid intermediate S26 (33mg, 63%).
IR(ATR):3361,2977,1730,1639cm-1.1HNMR (500MHz, acetone-d6):δ8.54-8.46(1H, M), 8.46-8.33 (1H, m, Ar-H), 8.15-8.07 (1H, m, Ar-H), 7.81 (1H, app.d, J=8.9Hz), 7.73- 7.62 (2H, m, Ar-H), 7.49 (1H, s), 7.36-7.15 (3H, m, Ar-H), 6.23 (1H, d, J=8.0Hz), 6.12 (1H, S), 6.03 (1H, d, J=8.6Hz), 4.81 (1H, app.t, J=7.8Hz, DABA1- α-CH), 4.73-4.62 (2H, m, Pal3-α-CH+Trp4-α-CH),4.46-4.36(2H,m,DABA1-β-CH+Leu3-α-CH),3.22-2.97(6H,m,NCH3 +Pal3-β-CH2+Trp4-β-CH2),2.82-2.74(1H,m,Pal3-β-CH2),1.78-1.69(1H,m,Leu3-γ-CH), 1.65(9H,s,3×CH3),1.62-1.56(1H,m,Leu3-β-CH2),1.50(1H,m,Leu3-β-CH2),1.36(9H,s,3 ×CH3),1.28(9H,s,3×CH3),1.24-1.14(3H,m,DABA1-γ-CH3),0.93-0.86(6H,m,2×Leu3- δ-CH3).LRMS[M+H+]880.5.HRMS(ESIm/z)[M+H+]C45H66N7O11Calculated value 880.4818, measured value 880.4824。
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- ((S) -2- ((uncle Butoxy carbonyl) amino)-N- methyl -3- (naphthalene -2- base) propionamido) ethyl) -5- isobutyl group 12,12- dimethyl -4,7,10- Three azepine tridecanoic acid (S27) of trioxy- -11- oxa- -3,6,8-
According to general step 6, in DMF (0.6mL), HATU (45mg, 120 μm of ol) and n,N-diisopropylethylamine are used The S25 (60 μm of ol) of resin-bonded and Boc-2-Nal-OH (38mg, 120 μm of ol) are coupled by (33 μ L, 180 μm of ol).? From resin crack (general step 7) after, by reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 minutes) it is pure Change peptide, obtains fluffy white solid intermediate S27 (31.4mg, 56%).
IR(ATR):3334,2979,1708,1660cm-1.1HNMR (400MHz, acetone-d6):δ8.10(1H,app.t,J =7.6Hz, Ar-H), 7.87-7.72 (4H, m, Ar-H), 7.67-7.58 (1H, m, Ar-H), 7.51-7.38 (4H, m, Ar-H), 7.29 (1H, ddd, J=8.4,7.3,1.4Hz, Ar-H), 7.23 (1H, app.td, J=7.5,1.2Hz, Ar-H), 6.18 (1H, D, J=8.1Hz), 6.11 (1H, d, J=12.7Hz), 5.98 (1H, d, J=8.8Hz), 5.12-5.04 (1H, m, DABA1- β- ), CH 4.94-4.84 (1H, m, DABA1- α-CH), 4.80 (1H, app.dt, J=9.1,4.6Hz, Nal2- α-CH), 4.72- 4.59 (1H, m, Trp4- α-CH), 4.50-4.32 (1H, m, Leu3- α-CH), 3.27 (1H, dd, J=14.1,4.0Hz, Nal2- β-CH2),3.13(3H,s,NCH3),3.11-3.01(2H,m,Trp4-β-CH2),2.98-2.89(1H,m,Nal2-β-CH2), 1.81-1.69(1H,m,Leu3-γ-CH),1.68-1.58(10H,m,3×CH3+Leu3-β-CH2),1.57-1.45(1H,m, Leu3-β-CH2),1.39-1.31(9H,m,3×CH3),1.27-1.08(12H,m,3×CH3+DABA1-γ-CH3),0.96- 0.85(6H,m,2×Leu3-δ-CH3).LRMS[M+H+]929.5.HRMS(ESIm/z)[M+Na+]C50H68N6O11Na calculated value 951.4838 measured value 951.4846.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- ((S) -2- ((uncle Butoxy carbonyl) amino) -3- cyclohexyl-N-methyl propionamido) ethyl) -5- isobutyl group -12,12- dimethyl -4,7,10- three Three azepine tridecanoic acid (S28) of oxo -11- oxa- -3,6,8-
In DMF (0.6mL), HATU (45mg, 120 μm of ol) and n,N-diisopropylethylamine (23 μ L, 126 μ are used Mol), the S25 of resin-bonded (60 μm of ol) and Boc-Cha-OH dicyclohexyl amine salt (54mg, 120 μm of ol) coupling reaction 16 is small When.From resin crack after (general step 7), by reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 points Clock) purified peptide, obtain fluffy white solid intermediate S28 (22mg, 41%).
IR(ATR):3346,2924,1728,1627cm-1.1HNMR(500MHz,CDCl3):δ7.99(1H,app.s,Ar- H),7.86(1H,s),7.63-7.53(1H,m,Ar-H),7.41(1H,s,Ar-H),7.28-7.20(2H,m,Ar-H),6.23 (1H,s),6.03(1H,s),5.64(1H,s),5.05-4.87(1H,m,DABA1-β-CH),4.79-4.66(1H,m,Trp4- α-CH),4.65-4.45(2H,m,DABA1-α-CH+Leu3-α-CH),4.45-4.15(1H,m,Cha2-α-CH),3.27- 3.07(2H,m,Trp4-β-CH2),2.98(3H,s,NCH3),1.97-1.81(1H,m,CH2),1.64(12H,m,3×CH3+ 0.5×CH2+Leu-β-CH2+CH),1.51-1.01(23H,m,6×CH3,CH2+CH+DABA1-γ-CH3),0.95-0.65 (7H,m,0.5×CH2+2×Leu3-δ-CH3).LRMS[M+H+]885.5.HRMS(ESIm/z)[M+Na+]C45H70N6O11Na meter Calculation value 907.5151, measured value 907.5150.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- ((S) -2- ((uncle Butoxy carbonyl) amino) -2- cyclohexyl-N-methyl propionamido) ethyl) -5- isobutyl group -12,12- dimethyl -4,7,10- three - 3,6,8 three azepine tridecanoic acid (S29) of oxygen -11- oxa-
In DMF (0.6mL), using HATU (124mg, 480 μm of ol), HOAt (324mg, 2.4 μm of mol) and N, N- bis- are different Propylethylamine (132 μ L, 720 μm of ol) is added in the S25 of resin-bonded, makes the S25 (60 μm of ol) and Boc- of resin-bonded Chg-OH (122mg, 480 μm of ol) coupling, reaction vessel vibrate 4.5 hours at room temperature.(the general step after being cracked from resin It is rapid 7), by reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 minutes) purified peptide, it is solid to obtain fluffy white Body intermediate S29 (19mg, 36%).
IR(ATR):3336,2977,1729,1641,1522cm-1.1HNMR (500MHz, acetone-d6):δ8.12(1H, App.d, J=8.1Hz, Ar-H), 7.72 (1H, d, J=8.9Hz, Ar-H), 7.71-7.65 (1H, m, Ar-H), 7.50 (1H, s, ), Ar-H 7.31 (1H, ddd, J=8.3,7.3,1.4Hz, Ar-H), 7.26 (1H, app.td, J=7.5,1.2Hz, Ar-H), 6.09 (1H, d, J=8.1Hz), 6.04 (1H, d, J=9.8Hz), 5.73 (1H, d, J=9.3Hz), 5.06-4.98 (1H, m, DABA1- β-CH), 4.73-4.69 (1H, m, DABA1- α-CH), 4.64 (1H, app.t, J=6.5Hz, Trp4- α-CH), 4.41-4.31(2H,m,Chg2-α-CH+Leu3-α-CH),3.23-3.06(2H,m,Trp4-β-CH2),3.01(3H,s, NCH3),1.75-1.70(1H,m,Leu3-γ-CH),1.67(9H,s,3×CH3),1.62-1.52(2H,m,Leu3-β-CH2+ CH2),1.52-1.44(2H,m,Leu3-β-CH2+CH2),1.32-1.14(7H,m,DABA1-γ-CH3+2×CH2),1.14- 1.02(3H,m),1.02-0.94(1H,m),0.95-0.87(6H,m,2×Leu3-δ-CH3).LRMS[M+H+]871.5.HRMS (ESIm/z)C45H70N6O11Na calculated value 893.4994, measured value 893.5001.
(2S, 3S) -2- (((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino) -3- (2- ((t-butoxy carbonyl)) ammonia Base)-N- methyl vinyl amido) methyl butyrate (S35)
According to general step 2, using n,N-diisopropylethylamine (500 μ L, 400 μm of ol) by amino acid 20 (219mg, 805 μm ol) it is loaded into CH2Cl2On 2- chlorine trityl chloride resin (514mg, 730 μm of ol) in (2mL).Then according to general step 5, allyl ammonia is sloughed using tetrakis triphenylphosphine palladium (0) (186mg, 161 μm of ol) and phenyl silane (2.0mL, 16.1mmol) Carbamate.In DMF (1.4mL), form method (general step 6) using the different peptide of solid phase, in HATU (554mg, 160 μm of ol) and Boc-Gly-OH (256mg, 1.46mmol) is set be coupled instead under n,N-diisopropylethylamine (381 μ L, 2.1mmol) effect It answers, obtains the different peptide S35 of resin-bonded.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methyl vinyl amido) ethyl) -5- ((R)-sec-butyl) -12,12- dimethyl -4,7,10- trioxy- -11- Three azepine tridecanoic acid (S36) of oxa- -3,6,8-
The different peptide S35 (60 μm of ol) of resin-bonded is deprotected into (general step 2) through Fmoc-, then according to general step 3, in DMF (0.6mL), using PyBOP (125mg, 240 μm of ol) and NMM (50 μ L, 480 μm of ol), with Fmoc-Ile-OH (85mg, 240 μm of ol) are coupled.Then according to general step 4, it is coupled with carbamate S12 (64mg, 120mol).? From resin crack (general step 7) after, by reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 minutes) it is pure Change peptide, obtains the depsipeptide S36 (17mg, 36%) of fluffy white solid.
IR(ATR):3308,2978,2929,1726,1637cm-1.1HNMR (500MHz, acetone-d6, main rotational isomeric Body): δ 8.12 (1H, d, J=7.9Hz), 7.80 (1H, app.d, J=8.9Hz, Ar-H), 7.69 (1H, d, J=7.3Hz), 7.59 (1H, app.d, J=8.6Hz, Ar-H), 7.51 (1H, s, Ar-H), 7.31 (1H, app.t, J=7.7Hz, Ar-H), 7.27 (1H, app.t, J=7.5Hz, Ar-H), 6.17-6.01 (2H, m), 5.85-5.75 (1H, m), 5.01-4.91 (1H, m, DABA1- β-CH), 4.75 (1H, app.t, J=7.2Hz, DABA1- α-CH), 4.64 (1H, app.t, J=7.3Hz, Trp4- α- CH),4.31-4.19(1H,m,Ile3-α-CH),3.92-3.83(1H,m,Gly2-α-CH2),3.75-3.68(1H,m,Gly2- α-CH2),3.20-3.07(2H,m,Trp4-β-CH2),2.85(3H,s,NCH3),1.87-1.72(1H,m,Ile3-β-CH), 1.67(9H,s,3×CH3),1.60-1.49(1H,m,Ile3-γ-CH2),1.44-1.33(18H,s,6×CH3),1.21(3H, D, J=6.9Hz, DABA1- γ-CH3),1.17-1.07(1H,m,Ile3-γ-CH2),0.96-0.76(6H,m,2×CH3) .LRMS[M+H+]789.4.HRMS(ESIm/z)[M+Na+]C39H60N6O11Na calculated value 811.4212, measured value 811.4227.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methyl vinyl amido) ethyl) -5- isopropyl -12,12- dimethyl -4,7,10- trioxy- -11- oxa- - Tri- azepine tridecane -1- of 3,6,8- is sour (S37)
The different peptide S35 (45 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc, then according to general step 6, In DMF (0.45mL), PyBOP (125mg, 240 μm of ol) and NMM (53 μ L, 480 μm of ol) and Fmoc-Val-OH are used (81.5mg, 240 μm of ol) are coupled.Then according to general step 4, it is coupled with carbamate S12 (63mg, 120 μm of ol). From resin crack (general step 7) after, by reversed-phase HPLC (50 to 100%MeCN, 40 minutes, 50%MeCN, 10 points Clock) peptide purified is obtained, obtain fluffy white solid depsipeptide S37 (19.7mg, 53%).
IR(ATR):3034,1735,1639,1384cm-1.1HNMR (400MHz, (acetone-d6, major rotomer): δ 8.14 (1H, d, J=8.3Hz, Ar-H), 7.70 (1H, d, J=7.4Hz, Ar-H), 7.52 (1H, s, Ar-H), 7.35-7.26 (2H,m,2xAr-H),6.21–6.13(2H,m,2xN-H),5.86(1H,m,N-H),5.04–4.97(1H,m,DABA1-β- CH),4.77(1H,m,DABA1-α-CH),4.69(1H,m,Trp4-α-CH),4.26(1H,m,Val3-α-CH),4.05–3.72 (2H,m,Gly2-α-CH2), 3.17 (2H, d, J=6.0Hz, Trp4- β-CH2),2.88(3H,s,NCH3),2.08(1H,m, Val3-β-CH),1.68(9H,s,3xCH3),1.42(18H,m,6xCH3), 1.24 (3H, d, J=7.0Hz, DABA1- γ- CH3), 0.95 (3H, d, J=6.7Hz, Val3- γ-CH3), 0.91 (3H, d, J=6.8Hz, Val3- γ-CH3).LRMS[M+H+] 775.5.HRMS(ESIm/z)[M+Na+]C38H58N6O11Na calculated value 797.4061, measured value 797.4063.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methyl vinyl amido) ethyl) oxa--3-12,12- dimethyl-4,7,10- trioxy--5- amyl-11-, Tri- azepine tridecanoic acid (S38) of 6,8-
The different peptide S35 (70 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc-, then according to general step 3, In DMF (0.7mL), PyBOP (144mg, 280 μm of ol) and NMM (58 μ L, 560 μm of ol) and Fmoc-Aha-OH are used (103mg, 280mol) is coupled.Then according to general step 4, it is coupled with carbamate S12 (75mg, 140 μm of ol).From After cracking (general step 7) on resin, pass through column chromatography (98:2v/vCH2Cl2: MeOH → 9:1v/v CH2Cl2: MeOH, The peptide 0.5vol%AcOH) is purified, white foam INTERMEDIATE S38 (30mg, 53%) is obtained.
IR(ATR):3356,2977,2932,1731,1638cm-1.1HNMR (500MHz, acetone-d6, main rotational isomeric Body): δ 8.12 (1H, d, J=8.2Hz, Ar-H), 7.69 (1H, d, J=7.6Hz, Ar-H), 7.60 (1H, d, J=8.5Hz), 7.52(1H,s,Ar-H),7.33-7.24(2H,m,Ar-H),6.19-6.01(1H,m),5.86-5.74(1H,m),5.04- 4.91 (1H, m, DABA1- β-CH), 4.80-4.68 (1H, m, DABA1- α-CH), 4.64 (1H, app.t, J=6.4Hz, Trp4- α-CH), 4.30 (1H, app.t, J=6.8Hz, Aha3- α-CH), 3.90-3.80 (1H, m, Gly2- α-CH2),3.79-3.66 (1H,m,Gly2-α-CH2),3.19-3.10(2H,m,Trp4-β-CH2),2.83(3H,s,NCH3), 1.76 (1H, dd, J= 16.3,8.3Hz,Aha3-β-CH2),1.67(9H,s,3×CH3),1.61-1.48(1H,m,Aha3-β-CH2),1.45-1.33 (20H,s,6×CH3+CH2),1.32-1.24(4H,m,2×CH2), 1.20 (3H, d, J=7.1Hz, DABA1- γ-CH3), 0.90-0.81(3H,m).LRMS[M+H+]803.5.HRMS(ESIm/z)[M+Na+]C40H62N6O11Na calculated value 825.4368, Measured value 825.4364.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methyl vinyl amido) ethyl) -5- cyclohexyl -12,12- dimethyl -4,7,10- trioxy- -11- oxa- - Tri- azepine tridecanoic acid (S39) of 3,6,8-
The different peptide S35 (60 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc-, according to general step 3, in DMF In (0.6mL), PyBOP (124mg, 240 μm of ol) and NMM (50 μ L, 480 μm of ol) and Fmoc-Chg-OH (91mg, 240 μ are used Mol it) is coupled.Then according to general step 4, in DMF (1mL), at n,N-diisopropylethylamine (21 μ L, 120 μm of ol) In the presence of, it is coupled carbamate S12 (64mg, 120 μm of ol).After cracking (general step 7) from resin, lead to Cross reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 minutes) purified peptide, obtain the contracting phenol of fluffy white solid Sour S39 (20mg, 40%).
IR(ATR):3308,2979,2929,1726,1637cm-1.1HNMR (400MHz, acetone-d6, main rotational isomeric Body): δ 8.11 (1H, app.d, J=8.1Hz, Ar-H), 7.74-7.60 (1H, m, Ar-H), 7.50 (1H, s, Ar-H), 7.36- 7.22(2H,m,Ar-H),6.25-6.12(2H,m),5.90-5.79(1H,m),5.06-4.91(1H,m,DABA1-β-CH), 4.81-4.70(1H,m,DABA1-α-CH),4.70-4.58(1H,m,Trp4-α-CH),4.29-4.19(1H,m,Chg3-α- CH),3.90-3.81(1H,m,Gly2-α-CH2),3.77-3.66(1H,m,Gly2-α-CH2),3.17-3.06(2H,m,Trp4- β-CH2),1.79-1.54(15H,3×CH3+2.5×CH2+Chg3-β-CH),1.43-1.32(18H,m,6×CH3),1.25- 0.93(8H,m,DABA1-γ-CH3+2.5×CH2).LRMS[M+H+]815.4.HRMS(ESIm/z)[M+H+]C41H62N6O11Meter Calculation value 815.4549, measured value 815.4548.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methyl vinyl amido) ethyl) -5- (cyclohexyl methyl) -12,12- dimethyl -4,7,10- trioxy- -11- Three azepine tridecanoic acid (S40) of oxa- -3,6,8-
The different peptide S35 (60 μm of ol) of resin-bonded is deprotected into (general step 2) through Fmoc-, according to general step 6, In DMF (0.6mL), HATU (45mg, 120 μm of ol) and n,N-diisopropylethylamine (33 μ L, 180 μm of ol) and Fmoc- are used Cha-OH (47mg, 120 μm of ol) is coupled.Then according to general step 4, with carbamate S12 (64mg, 120 μm of ol) Coupling.From resin crack after (general step 7), by reversed-phase HPLC (50-100%MeCN, 60 minutes；50%MeCN, 10 points Clock) purified peptide, obtain fluffy white solid depsipeptide S40 (17mg, 34%).
IR(ATR):3390,2977,2936,1730,1644cm-1.1HNMR (500MHz, acetone-d6, rotational isomer ratio Example 4:1, major rotomer): δ 8.12 (1H, app.d, J=8.2Hz, Ar-H), 7.72-7.66 (1H, m), 7.61 (1H, App.d, J=8.9Hz, Ar-H), 7.52 (1H, s, Ar-H), 7.32 (1H, ddd, J=8.3,7.1,1.4Hz, Ar-H), 7.27 (1H, app.td, J=7.5,1.2Hz, Ar-H), 6.16-5.97 (2H, m), 5.89-5.69 (1H, m), 5.00-4.93 (1H, m, DABA1-β-CH),4.79-4.70(1H,m,DABA1-α-CH),4.68-4.60(1H,m,Trp4-α-CH),4.42-4.35 (1H,m,Cha3-α-CH),3.96(1H,t,J5.5Hz),3.87-3.81(1H,m,Gly2-α-CH2),3.75-3.69(1H,m, Gly2-α-CH2),3.19-3.11(2H,m,Trp4-β-CH2),2.82(3H,s,NCH3),1.84-1.75(1H,m,CH2), 1.67-1.56(13H,m,3×CH3+1.5×CH2+Cha3-β-CH2),1.48-1.31(22H,m,6×CH3+Cha3-β-CH2+ Cha3-γ-CH+CH2),1.24-1.08(5H,m,DABA1-γ-CH3+CH2),1.00-0.77(2H,m,CH2).LRMS[M+H+] 829.5。HRMS(ESIm/z)[M+Na+]C42H64N6O11Na calculated value 851.5425, measured value 851.4526.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methyl vinyl amido) ethyl) -5- ((S) -1- ethoxy) -12,12- dimethyl -4,7,10- trioxy- - Three azepine tridecane -1- of 11- oxa- -3,6,8- is sour (S41)
The different peptide S35 (40 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc, according to general step 6, in DMF In (0.4mL), using HATU (30.4mg, 80 μm of ol) and n,N-diisopropylethylamine (20 μ L, 120 μm of ol), dual coupling is extremely Fmoc-Thr-OH (13.7mg, 80 μm of ol).Carbamate S12 (46mg, 90 μm of ol) are then coupled according to general step.? After cracking (general step 7) from resin, by reversed-phase HPLC (50 to 100%MeCN, 40 minutes) purified peptide, obtain fluffy White solid depsipeptide S41 (18.2mg, 49%).
IR(ATR):3004,1709,1422,1359cm-1.1HNMR (400MHz, acetone-d6, major rotomer): δ 8.08 (1H, d, J=8.1Hz, Ar-H), 7.66 (1H, d, J=8.6Hz, Ar-H), 7.49 (1H, s, Ar-H), 7.30-7.21 (2H,m,2xAr-H),6.37(1H,m,N-H),6.24(1H,m,N-H),5.85(1H,m,N-H),4.97(1H,m,DABA1-β- CH),4.69(1H,m,DABA1-α-CH),4.63(1H,m,Trp4-α-CH),4.24(2H,m,Thr3-α-CH+Thr3-β- CH),3.92–3.77(2H,m,Gly2-α-CH2),3.14(2H,m,Trp4-β-CH2),2.80(3H,s,NCH3),1.63(9H, s,3xCH3),1.38–1.34(18H,m,6xCH3), 1.18 (3H, d, J=7.1Hz, DABA1- γ-CH3), 1.08 (3H, d, J= 6.4Hz,Thr3-γ-CH3).LRMS[M+H+]777.5.HRMS(ESIm/z)[M+Na+]C37H56N6O12Na calculated value 799.3853 measured value 799.3850.
(2S, 5S, 9S) -5- (2- (tertbutyloxycarbonyl) -2- oxoethyl) -9- ((1- (tertbutyloxycarbonyl) -1-H- indoles - 3- yl) methyl) -2- ((S) -1- (2- ((tertbutyloxycarbonyl) amino)-N- methyl vinyl amido) ethyl) -12,12- dimethyl 4, Three azepine tridecane -1- of 7,10- trioxy- -11- oxa- -3,6,8- is sour (S42)
The different peptide S35 (60 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc-, according to general step 6, in DMF In (0.6mL), Fmoc-Asp (O is coupled to using PyBOP (125mg, 240 μm of ol) and NMM (53 μ L, 480 μm of ol)tBu)-OH (99mg, 240 μm of ol).Then according to general step 4, carbamate S12 (62mg, 120 μm of ol) is coupled.From tree After cracking on rouge (general step 7), pass through column chromatography eluting peptide (eluent 95:5v/v CH2Cl2: MeOH, 0.1 volume % AcOH), yellow oily depsipeptide S42 (37.2mg, 69%) is obtained.
IR(ATR):3359,2978,2933,1726,1640cm-1.1HNMR (300MHz, acetone-d6, main rotational isomeric Body): δ 8.11 (1H, d, J=7.8Hz, Ar-H), 7.79 (1H, d, J=6.9Hz, Ar-H), 7.52 (1H, s, Ar-H), 7.34- 7.25(2H,m,2xAr-H),6.36-6.24(2H,m,2xN-H),5.83(1H,m,N-H),4.93(1H,m,DABA1-β-CH), 4.74-4.63(3H,m,DABA1-α-CH+Asp3-α-CH+Trp4-α-CH),3.94–3.66(2H,m,Gly2-α-CH2), 3.17 (2H, d, J=6.3Hz, Trp4- β-CH2),2.75(3H,s,NCH3),2.67–2.60(2H,m,Asp3-β-CH2),1.66 (9H,m,3xCH3),1.41–1.38(27H,m,9xCH3), 1.10 (3H, d, J=6.9Hz, DABA1- γ-CH3).LRMS[M+H+]847.6.HRMS(ESI m/z)[M+Na+] calculated value C41H62N6O13Na869.4272, measured value 869.4272.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methylacetamido) ethyl)-5- (4- ((tertbutyloxycarbonyl) amino) butyl)-12,12-dimethyl-4, Three azepine tridecane -1- of 7,10- trioxy- -11- oxa- -3,6,8- is sour (S43)
The different peptide S35 (45 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc, according to general step 6, in DMF In (0.6mL), PyBOP (125mg, 240 μm of ol) and NMM (53 μ L, 480 μm of ol) and Fmoc-Lys (Boc)-OH are used (112mg, 240 μm of ol) coupling.Carbamate S12 (63mg, 126 μm of ol) are then coupled according to general step 4.From resin After upper cracking (general step 6), pass through column chromatography (97:3v/v CH2Cl2: i-PrOH, 0.1vol% acetic acid -9:1v/v CH2Cl2: i-PrOH, 0.1% acetic acid) obtain white foam depsipeptide S43 (15.4mg, 38%).
IR(ATR):3325,2977,1701,1647cm-1.1HNMR (400MHz, acetone-d6, major rotomer): δ 8.08 (1H, d, J=10.4Hz, Ar-H), 7.66 (1H, d, J=9.2Hz, Ar-H), 7.47 (1H, s, Ar-H), 7.30-7.20 (2H,m,2xAr-H),6.09(2H,m,2xN-H),5.88(1H,m,N-H),4.96(1H,m,DABA1-β-CH),4.72-4.61 (2H,m,DABA1-α-CH+Trp4-α-CH),4.30(1H,m,Lys3-α-CH),3.94–3.66(2H,m,Gly2-α-CH2), 3.12 (2H, d, J=8.0Hz, Trp4- β-CH2), 2.99 (2H, d, J=8.4Hz, Lys3- β-CH2),2.80(3H,s,NCH3), 1.63(6H,m,Lys-γ-CH2+Lys-δ-CH2+Lys-ε-CH2),1.37–1.24(36H,m,12xCH3),1.16(3H,d,J =9.2Hz, DABA1- γ-CH3).LRMS[M+H+]904.6.HRMS(ESIm/z)[M+Na+]C44H69N7O13Na calculated value 926.4851 measured value 926.4854.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) Amino-N-methyl acetylamino) ethyl) -12,12- dimethyl -4,7,10- trioxy- -5- (4- (trifluoromethyl) benzyl Base) 6 miscellaneous three azepine tridecanoic acid (S45) of -3,6,8- of -11- oxygen
The different peptide S35 (88mol) of resin-bonded deprotects (general step 2) through Fmoc-, according to general step 3, in DMF In (0.9mL), PyBOP (183mg, 352 μm of ol) and NMM (73 μ L, 704 μm of ol) and Fmoc-Phe (4-CF are used3)-OH (160mg, 352 μm of ol) coupling.Then according to general step 4, carbamate S12 (94mg, 176mol) is coupled.? From resin crack after (general step 7), by reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 minutes) it is pure Change peptide, obtains fluffy white solid depsipeptide S45 (29mg, 36%).
IR(ATR):3374,2979,2932,1731,1666cm-1.1HNMR(400MHz,CDCl3, main rotational isomeric Body): δ 8.04 (1H, app.d, J=8.2Hz, Ar-H), 7.76-7.60 (1H, m), 7.64 (1H, d, J=6.8Hz), 7.53 (1H, app.d, J=7.5Hz, Ar-H), 7.47-7.36 (4H, m, Ar-H), 7.30-7.14 (3H, m, Ar-H), 5.93 (1H, s),5.82(1H,s),5.54(1H,s),4.98-4.82(1H,m,DABA1-β-CH),4.76-4.32(3H,m,DABA1-α-CH +4-CF3-Phe3-α-CH+Trp4-α-CH),3.85-3.67(2H,m,Gly2-α-CH2),3.17-3.05(3H,m,4-CF3- Phe3-β-CH2+Trp4-β-CH2),3.00-2.91(1H,m,Trp4-β-CH2),2.73(3H,s,NCH3),1.62(9H,s,3 ×CH3),1.42-1.19(18H,m,6×CH3), 1.15 (3H, d, J=6.9Hz, DABA1- γ-CH3).LRMS[M+H+] 891.4.HRMS(ESIm/z)[M+H+] calculated value C43H57F3N6O11891.4110 measured value 891.4103.
(2S, 5S, 9S) -5- (4- (tertbutyloxycarbonyl) benzyl) -9- ((1- (tertbutyloxycarbonyl) -1H indol-3-yl) first Base) -2- ((S) -1- (2- ((tertbutyloxycarbonyl) amino)-N- methylacetamido) ethyl) -12,12- dimethyl 4,7,10- tri- Three azepine tridecane -1- of oxygen -11- oxa- -3,6,8- is sour (S46)
The different peptide S35 (60 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc-, according to general step 6, in DMF In (0.6mL), PyBOP (125mg, 240 μm of ol) and NMM (53 μ L, 480 μm of ol) and Fmoc-Tyr (O are usedtBu)-OH (110mg, 240 μm of ol) coupling.Then according to general step 4, it is coupled with carbamate S12 (63mg, 120 μm of ol).? After cracking from resin (general step 7), pass through column chromatography eluting peptide (eluent: 95:5v/vCH2Cl2: MeOH, 0.1 body Product % acetic acid), obtain yellow oily depsipeptide S46 (34.8mg, 56%).
IR(ATR):3350,2978,2929,1726,1637cm-1.1HNMR(400MHz,CDCl3, main rotational isomeric Body): δ 8.06 (1H, d, J=7.2Hz, Ar-H), 7.55 (1H, d, J=7.6Hz, Ar-H), 7.39 (1H, s, Ar-H), 7.29- 7.18 (2H, m, 2xAr-H), 7.04 (2H, d, J=8.4Hz, 2xAr-H), 6.82 (2H, d, J=8.4Hz, 2xAr-H), 5.72- 5.61(2H,m,2xN-H),4.84(1H,m,DABA1-β-CH),4.65(1H,m,DABA1-α-CH),4.57(1H,m,Trp4- α-CH),4.52(2H,m,Tyr3-α-CH),3.91–3.71(2H,m,Gly2-α-CH2),3.12(2H,m,Trp4-β-CH2), 2.98(2H,m,Trp4-β-CH2),2.79(3H,s,NCH3),1.63(9H,s,3xCH3),1.40–1.25(27H,m,9xCH3), 1.14 (3H, d, J=6.8Hz, DABA1- γ-CH3).LRMS[M+H+]895.5.HRMS(ESIm/z)[M+Na+] C46H66N6O12Na calculated value 895.4817, measured value 895.4810.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methylacetamido) ethyl) -12,12- dimethyl -4,7,10- trioxy- -5- (pyridin-3-yl (first Base) three azepine tridecanoic acid (S47) of -11- oxa- -3,6,8-
The different peptide S35 (60 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc-, according to general step 3, in DMF In (0.6mL), Fmoc-3-Pal-OH is coupled to using PyBOP (125mg, 240 μm of ol) and NMM (50 μ L, 480 μm of ol) (92mg, 240 μm of ol).Then according to general step 4, carbamate S12 (64mg, 120 μm of ol) is coupled.From tree On rouge crack (general step 7) after, by reversed-phase HPLC (50-100%MeCN, 60 minutes；50%MeCN, 10 minutes) purifying Peptide obtains the depside S47 (14mg, 29%) of fluffy white solid.
IR(ATR):3366,2979,2936,1726,1660cm-1.1HNMR (500MHz, acetone-d6, main rotational isomeric Body): δ 8.70-8.61 (2H, m, NH+Ar-H), 8.21 (1H, app.d, J=7.9Hz, Ar-H), 8.11 (1H, app.d, J= 8.2Hz, Ar-H), 7.98 (1H, d, J=8.8Hz, Ar-H), 7.83-7.81 (1H, m, Ar-H), 7.64 (1H, d, J=7.0Hz, ), Ar-H 7.50 (1H, s, Ar-H), 7.33-7.28 (1H, m, Ar-H), 7.25 (1H, app.td, J=7.5,1.2Hz, Ar-H), 6.37-6.21(2H,m,2×NH),5.86-5.73(1H,m),4.97-4.89(1H,m,DABA1-β-CH),4.81-4.73 (1H, m, J=7.0Hz, 3-Pal3- α-CH), 4.71-4.62 (1H, m, DABA1- α-CH), 4.61-4.52 (1H, m, Trp4- α- CH),3.94-3.81(1H,m,Gly2-α-CH2),3.78-3.69(1H,m,Gly2-α-CH2), 3.31 (1H, app.dt, J= 15.6,7.6Hz,3-Pal3-β-CH2),3.20-3.06(3H,m,3-Pal3-β-CH2+Trp4-β-CH2),2.83(3H,s, NCH3),1.65(9H,s,3×CH3),1.43-1.31(18H,s,6×CH3), 1.18 (3H, d, J=7.1Hz, DABA1- γ- CH3).LRMS[M+H+]829.5.HRMS(ESIm/z)[M+H+] calculated value C41H57N7O11824.4188 measured value 824.4182.
Bis- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (the 2- ((uncles of (2S, 5S, 9S) -5,9- Butoxy carbonyl) amino)-N- methylacetamido) ethyl) -12,12- dimethyl -4,7,10- trioxy- -11- oxa- 3,6,8- Three azepine tridecane -1- are sour (S48)
The different peptide S35 (60 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc, according to general step 7, in DMF In (0.6mL), Fmoc-Trp (Boc)-OH is coupled to using PyBOP (125mg, 240 μm of ol) and NMM (53 μ L, 480 μm of ol) On (126.4mg, 240 μm of ol).Then according to general step 4, carbamate S12 (90.4mg, 172 μm of ol) are carried out even Connection.After cracking from resin (general step 7), pass through column chromatography eluting peptide (eluent: 95:5v/v CH2Cl2: it is inactivating Silica on MeOH, 0.1vol% acetic acid), obtain yellow oily depsipeptide S48 (30.2mg, 52%).
IR(ATR):3352,2980,2928,1731,1646,1553cm-1.1HNMR(500MHz,CDCl3, main to rotate Isomers): δ 8.04 (1H, m, 2xAr-H), 7.53 (1H, d, J=7.6Hz, 2xAr-H), 7.40 (2H, m, Ar-H), 7.28- 7.11(4H,m,4xAr-H),6.29(1H,m,N-H),6.06(1H,m,N-H),5.57(1H,m,N-H),4.84(1H,m, DABA1-β-CH),4.68-4.52(2H,m,Trp3-α-CH+Trp4-α-CH),4.47(1H,m,DABA1-α-CH),3.75– 3.48(2H,m,Gly2-α-CH2),3.16-2.91(4H,m,Trp3-β-CH2+Trp4-β-CH2),2.88(3H,s,NCH3), 1.61(18H,s,6xCH3),1.42–1.22(18H,m,6xCH3), 1.11 (3H, d, J=6.6Hz, DABA1- γ-CH3).LRMS [M+H+]962.6.HRMS(ESIm/z)[M+H+]C49H68N7O11Calculated value 962.4876, measured value 962.4869.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methylacetamido) ethyl) oxa--3-12,12- dimethyl-4,7,10- trioxy--5- phenyl-11-, Tri- azepine tridecane -1- of 6,8- is sour (S49)
The different peptide S35 (70 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc, according to general step 6, in DMF In (0.7mL), HOAt (11.2mg, 84 μm of ol) and DIC (13.3 μ L, 84 μm of ol) and Fmoc-Phg-OH (31.5mg, 84 μ are used Mol it) is coupled.Then according to general step 4, carbamate S12 (73.6mg, 140 μm of ol) is coupled.From resin After cracking (general step 7), the mixture peptide of 9:1 diastereoisomer is obtained, reversed-phase HPLC (50 to 100% can be passed through MeCN, 40 minutes) it is easily separated, obtain the fluffy white solid depsipeptide S49 of single diastereoisomer (14.5mg, 25%).
IR(ATR):3343,2978,2933,1727,1700,1638cm-1.1HNMR (400MHz, acetone-d6, main to revolve Turn isomers): δ 8.13 (1H, d, J=7.6Hz, Ar-H), 7.72 (1H, d, J=6.8Hz, Ar-H), 7.52 (1H, s, Ar-H), 7.46(2H,d,J7.2Hz,2xAr-H),7.47-7.24(5H,m,5xAr-H),6.66(1H,m,N-H),6.27(1H,m,N- H),5.80(1H,m,N-H),5.57(1H,m,Phg3-α-CH),4.99(1H,m,DABA1-β-CH),4.79(1H,m,DABA1- α-CH),4.65(1H,m,Trp4-α-CH),3.95-3.71(2H,m,Gly2-α-CH2),3.17(2H,m,Trp4-β-CH2), 2.88(3H,s,NCH3),1.67(9H,s,3xCH3),1.40(9H,m,3xCH3),1.35(9H,m,3xCH3),1.25(3H,d,J =6.8Hz, DABA1- γ-CH3).LRMS[M+H+]809.5.HRMS(ESIm/z)[M+Na+]C41H56N6O11Na calculated value 809.4085 measured value 809.4080.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methylacetamido) ethyl) -12,12- dimethyl -4,7,10- trioxy- -5- phenethyl -11- oxa- - Tri- azepine tridecane -1- of 3,6,8- is sour (S50)
The different peptide S35 (70 μm of ol) of resin-bonded deprotects (general step 2) through Fmoc, according to general step 6, in DMF In (0.7mL), using HOAt (11.2mg, 84 μm of ol) and DIC (13.3 μ L, 84 μm of ol) and Fmoc-HPhe-OH (33.7mg, 84 μm ol) coupling.Then according to general step 4, carbamate S12 (73.6mg, 140 μm of ol) is coupled.From resin After upper cracking (general step 7), the mixture peptide of 9:1 diastereoisomer is obtained, reversed-phase HPLC (50 to 100% can be passed through MeCN, 40 minutes) it is easily separated, it obtains as single diastereoisomer fluffy white solid depsipeptide S50 (18.8mg, 27%).
IR(ATR):3342,2977,2931,1730cm-1.1HNMR(500MHz,CDCl3, major rotomer): δ 8.11 (1H, d, J=8.0Hz, Ar-H), 7.69 (1H, d, J=7.5Hz, Ar-H), 7.53 (1H, s, Ar-H), 7.32-7.12 (7H,m,7xAr-H),6.39(1H,m,N-H),6.25(1H,m,N-H),5.95(1H,m,N-H),4.96(1H,m,DABA1-β- CH),4.71(1H,m,DABA1-α-CH),4.64(1H,m,Trp4-α-CH),4.36(1H,m,HPhe3-α-CH),3.96– 3.69(2H,m,Gly2-α-CH2),3.15(2H,m,Trp4-β-CH2),2.80(3H,s,NCH3),2.64(2H,m,HPhe3-β- CH2),2.02(2H,m,HPhe3-γ-CH2),1.63(9H,s,3xCH3),1.36–1.34(18H,m,6xCH3),1.16(3H, D, J=7.1Hz, DABA1- γ-CH3).LRMS[M+H+]837.5.HRMS(ESIm/z)[M+Na+]C43H60N6O11Na calculated value 859.4218 measured value 859.4205.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- (2- ((tertiary fourth oxygen Carbonyl) amino)-N- methylacetamido) ethyl) -12,12- dimethyl -5- (naphthalene -2- ylmethyl) -4,7,10- trioxy- - Three azepine tridecanoic acid (S51) of 11- oxa- -3,6,8-
The different peptide S35 (60 μm of ol) of resin-bonded is through Fmoc- de-protected (general program 2), according to general step 3, In DMF (0.6mL), Fmoc-2-Nal-OH is coupled to using PyBOP (124mg, 240 μm of ol) and NMM (50 μ L, 480 μm of ol) (104mg, 240 μm of ol).It is right in the presence of DIPEA (21 μ L, 120 μm of ol) in DMF (1mL) then according to general step 4 Carbamate S12 (64mg, 120mol) is coupled.After cracking from resin (general step 7), pass through reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 minutes) purified peptide, obtain the depsipeptide S51 of fluffy white solid (22mg, 42%).
IR(ATR):3390,2978,2936,1730,1644cm-1.1HNMR (500MHz, acetone-d6, it is main rotation it is different Structure body): δ 8.11 (1H, d, J=8.2Hz, Ar-H), 7.86-7.61 (6H, m, 5 × Ar-H+NH), 7.50 (1H, s, Ar-H), 7.45-7.37(3H,m,Ar-H),7.34-7.21(2H,m,Ar-H),6.22-6.12(2H,m,NH),5.82(1H,s,NH), 5.01-4.92(1H,m,DABA1-β-CH),4.82-4.69(2H,m,DABA1-α-CH+2-Nal3-α-CH),4.66-4.58 (1H,m,Trp4-α-CH),3.93-3.83(1H,m,Gly2-α-CH2), 3.72 (1H, dd, J=17.0,4.2Hz, Gly2- α- CH2),3.39-3.25(1H,m,2-Nal3-β-CH2),3.19-3.05(3H,m,2-Nal3-β-CH2+Trp4-β-CH2),2.84 (3H,s,NCH3),1.65(9H,s,3×CH3),1.46-1.30(18H,m,6×CH3), 1.17 (3H, d, J=7.2Hz, DABA1-γ-CH3).LRMS[M+H+]873.4.HRMS(ESIm/z)[M+Na+]C46H60N6O11Na calculated value 895.4212 is surveyed Magnitude 895.4212.
(2S, 5S, 9S) -9- ((1- (tertbutyloxycarbonyl) -1H- indol-3-yl) methyl) -2- ((S) -1- ((S) -2- ((uncle Butoxy carbonyl) amino) -3- (3- ((t-butyldimethylsilyi) oxygroup) phenyl)-N- methyl propanamide base) ethyl) -5- Three azepine tridecanoic acid (S52) of (cyclohexyl methyl) -12,12- dimethyl -4,7,10- trioxa -11- oxa- -3,6,8-
According to general step 6, in DMF (0.9mL), HATU (65mg, 174 μm of ol) and n,N-diisopropylethylamine are used The amino acid S25 (87 μm of ol) of resin-bonded and Fmoc-Cha-OH (68mg, 174 μm of ol) are coupled by (48 μ L, 260 μm of ol). Then according to general step 4, carbamate S12 (93mg, 174 μm of ol) is coupled.Then according to general step 5, make With tetrakis triphenylphosphine palladium (0) (20mg, 17 μm of ol) and phenyl silane (20 μ L, 170 μm of ol), in CH2Cl2In (1mL), remove Allyl carbamate.Then, the solution in DMF (0.87mL) uses HATU (49mg, 130 μm of ol), HOAt (25mg, 130 μ Mol) and n,N-diisopropylethylamine (23 μ L, 130 μm of ol) is coupled to Boc-m-Tyr (OTBS)-OH (51mg, 130 μm of ol).? From resin crack (general step 7) after, by reversed-phase HPLC (50-100%MeCN, 40 minutes；50%MeCN, 10 minutes) it is pure Change peptide, obtains the depsipeptide S52 (18mg, 20%) of fluffy white solid.
IR(ATR):3314,2928,2856,1730,1630cm-1.1HNMR(500MHz,CDCl3, main rotational isomeric Body): δ 8.08-7.92 (1H, m, Ar-H), 7.79 (1H, s), 7.61-7.52 (1H, m, Ar-H), 7.45-7.36 (1H, m, Ar- H),7.31-7.15(2H,m,Ar-H),7.01-6.87(1H,m,Ar-H),6.72-6.46(3H,m,Ar-H),6.11-5.73 (1H,m,NH),5.69-5.53(1H,m,NH),4.94-4.84(1H,m,DABA1-β-CH),4.76-4.60(3H,m,DABA1- α-CH+m-Tyr2-α-CH+Trp4-α-CH),4.46-4.19(1H,m,Cha3-α-CH),3.30-3.07(2H,m,Trp4-β- CH2),3.02(3H,s,NCH3),2.98-2.87(1H,m,m-Tyr2-β-CH2),2.61-2.46(1H,m,m-Tyr2-β- CH2),1.78-0.74(58H,m).LRMS[M+H+]1049.4.HRMS(ESIm/z)[M+H+]C55H84N6O12Si calculated value 1049.5989, measured value 1049.5981.
The synthesis of analog
General step 8: pass through fragment condensation strategy synthetic analogues
Condition A:
To the CH of amine 18 (15-25mol, 1.2eq.)2Cl2With DMF (1:1v/vCH2Cl2: DMF0.1M) solution in be added Depsipeptide S24, S26, S27, S29 and S53 (13-21 μm of ol, 1eq.) and HOAt (5-5.5eq.), are added DIC immediately after (1eq.).Reactant is stirred at room temperature 1.5-2 hours, at this point, by reaction mixture H2O (5mL) and saturation NaHCO3Water Solution (5mL) dilution, and extracted with EtOAc (30mL).Organic phase another part is saturated NaHCO3Aqueous solution (5mL) is washed It washs, through anhydrous MgSO4It dries and is concentrated in vacuo, obtain thick residue, be then resuspended in TFA and contain CH2Cl2's iPr3(1:1v/v TFA:CH in the mixture of SiH2Cl2, 2.5vol%iPr3SiH, 0.1mL/ μm of ol), and will react in room temperature Lower stirring (1.5-16 hours).Solvent is removed in vacuum, obtains thick residue, it is purified by reversed-phase HPLC.
Condition B:
To the 1:1v/vCH of EDCHCl (12-30 μm of ol, 1.3-3.3eq.)2Cl2: DMF (180-240 μ L) solution is added NMM (19-30 μm of ol, 1.3eq.), and solution is vibrated at room temperature 15 minutes.Then the solution is added drop-wise to depsipeptide S28, S36-S43, S45-S52 (15-25 μm of ol, 1eq.), amine 18 (28-50 μm of ol, 1.2-3.3eq.) and HOAt (70-120 μ Mol, 5eq.) solution in.And reaction is stirred at room temperature 3-4 hours.At this point, by reaction mixture EtOAc (15- 25mL) dilute, and use HCl (0.5M, 4-7mL), saturation NaHCO3Aqueous solution (5 × 4-7mL), H2O (4-7mL) and salt water (4- It 7mL) washs, uses MgSO4It is dry.Then solvent is removed in a vacuum, obtains thick residue, is re-dissolved in CH2Cl2, TFA and iPr3SiH (1:1CH2Cl2: TFA, 2.5vol.%iPr3SiH, 0.1mL/ μm of ol) mixture in, and will react at room temperature It stirs (5-16 hours).Solvent is removed in vacuum, obtains thick residue, it is purified by reversed-phase HPLC.
(((S) -1- (((2S, 3S) -3- ((S) -2- Amino-N-methyl -3- (pyridin-3-yl) propionamido) -1- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) Amino)-1- oxo-butanes-2- base) amino)-4- methyl-1-oxo-pentane-2- base) carbamoyl)-L-Trp (8)
According to the condition A of general step 8, at room temperature, HOAt (18mg, 134 μm of ol) and DIC (4.3 μ L, 27 μm of ol) are deposited Under, make depside S26 (24mg, 27 μm of ol) and amine 18 (7.2mg, 32 μm of ol) in CH2Cl2: in DMF (1:1v/v, 260 μ L) Reaction 1.5 hours obtains protection analog completely.According to universal method 8, the compound is with TFA and contains iPr3SiH's CH2Cl2Mixture handles (1:1v/vTFA:CH2Cl2, 2.5vol.%iPr3SiH, 2.7mL), in reversed-phase HPLC (0 to 50% MeCN, 40 minutes；10 minutes, 100%H2O) after purification, obtain amorphous white solid 8 (as formates) (11mg, 2 steps, Yield 46%).
IR(ATR):2951,2836,1662cm-1.1HNMR(500MHz,CD3OD, the ratio 1.3:1 of rotational isomer, rotation Turn isomers 1): δ 8.69-8.64 (1H, m, NH), 8.63-8.57 (1H, m, NH), 8.57-8.49 (1H, m, NH), 8.12 (1H, D, J=7.5Hz, Ar-H), 7.98 (1H, d, J=8.0Hz, Ar-H), 7.66-7.59 (2H, m, Ar-H+H-6), 7.58-7.51 (1H,m,Ar-H),7.35-7.23(1H,m,Ar-H),7.11(1H,s,Ar-H),7.10-7.04(1H,m,Ar-H),7.03- 6.93 (1H, m, Ar-H), 5.74 (1H, d, J=2.7Hz, H-1 '), 5.69 (1H, d, J=8.0Hz, H-5), 5.00-4.91 (1H,m,Pal2-α-CH),4.89-4.81(1H,m,DABA1-β-CH),4.76-4.72(1H,m,DABA1-α-CH),4.64- 4.37(3H,m,H-2’+H-4’+Trp4-α-CH),4.30-4.04(2H,m,Leu3-α-CH),3.52-3.41(1H,m,H- 5’),3.30-3.24(2H,m,H-5’+Trp4-β-CH2),3.23-3.11(3H,m,Pal2-β-CH2+Trp4-β-CH2),2.81 (3H,s,NCH3), 2.31 (1H, dt, J=13.8,6.9Hz, H-2 '), 2.23 (1H, ddd, J=13.7,7.5,6.2Hz, H- 2’),1.86-1.77(1H,m,H-2’),1.78-1.61(3H,m,H-2’+Leu3-γ-CH),1.60-1.40(2H,m,Leu3- β-CH2), 1.28 (3H, d, J=7.1Hz, DABA1- γ-CH3), 1.24 (3H, d, J=7.1Hz, DABA1- γ-CH3),0.98- 0.86(6H,m,2×Leu3-δ-CH3).13CNMR(125MHz,CD3OD, the ratio 1.3:1 of rotational isomer, rotational isomer 1): δ 176.0 (C=O), 175.6 (C=O), 171.3 (C=O), 169.2 (C=O), 166.1 (C=O), 159.8 (C=O), 151.9 (C=O), 147.6,144.4,142.0,141.6,137.9,128.9,126.5,124.6 (× 2), 122.2,119.6, 119.1,111.9,110.6,102.2,94.9,80.6,76.2,56.4,54.7,54.3,53.0,52.3,44.6,41.6, 35.6,34.8,28.8,28.4,25.4,22.4,12.5.1HNMR(500MHz,CD3OD, the ratio 1.3:1 of rotational isomer, rotation Turn isomers 2): δ 8.06 (1H, d, J=8.0Hz, Ar-H), 7.48 (1H, d, J=8.1Hz, H-6), 5.77 (1H, d, J= 3.0Hz, H-1 '), 5.64 (1H, d, J=8.1Hz, H-5), 5.11-5.04 (1H, m, DABA1- β-CH), 4.64-4.37 (2H, m,DABA1-α-CH+Pal2-α-CH),3.40-3.32(1H,m,Pal2-β-CH2),3.11-3.05(1H,m,Pal2-β-CH2), 3.01(3H,s,NCH3)；13C NMR(125MHz,CD3OD, the ratio 1.3:1 of rotational isomer, rotational isomer 2): δ 165.7 (C=O), 146.7,141.7,51.9,33.4,30.6,12.2.LRMS [M+H+]833.4.HRMS(ESIm/z)[M+H+] C40H52N10O10Calculated value 833.3940, measured value 833.3938.
(((S) -1- (((2S, 3S) -3- ((S) -2- Amino-N-methyl -3- (naphthalene -2- base) propionamido) -1- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- Oxo-butanes-2- base) amino)-4- methyl-1-oxo-pentane-2- base) carbamoyl)-L-Trp (9)
According to the condition A of general step 8, at room temperature, HOAt (23mg, 165 μm of ol) and DIC (5.3 μ L, 33 μm of ol) are deposited Under, make depsipeptide S27 (30mg, 33 μm of ol) and amine 18 (9.8mg, 40 μm of ol) in CH2Cl2Middle reaction 2.5 hours, obtains Protection analog completely.According to universal method 8, the compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture processing (1: 1v/vTFA:CH2Cl2, 2.5vol.%iPr3SiH, 3.3mL), and reversed-phase HPLC (0 to 50%MeCN, 40 minutes；10 minutes, 100%H2O amorphous white solid 9 (formates) (10mg, 2 steps, 32%)) are obtained after purification.
IR(ATR):3264,3056,2954,2918,2851,1672,1636cm-1.1HNMR(500MHz,DMSO-d6, rotation Turn the ratio 1.8:1 of isomers, main rotational isomer): 10.78 (1H, s), 8.52 (1H, s), 8.32 (1H, s), 8.20 (2H, s), 7.88-7.77 (3H, m, Ar-H), 7.65 (1H, app.s, Ar-H), 7.58 (1H, d, J=8.1Hz, H-6), 7.54- 7.39(1H,m,Ar-H),7.47-7.35(3H,m,Ar-H),7.32-7.27(1H,m,Ar-H),7.12-7.07(1H,m,Ar- ), H 7.05-6.99 (1H, m, Ar-H), 6.97-6.88 (1H, m, Ar-H), 6.46 (1H, d, J=8.0Hz), 6.39 (1H, d, J =8.0Hz), 6.13 (1H, s), 5.64 (1H, d, H-1 '), 5.58 (1H, d, H-5), 4.46 (1H, app.t, J=9.2Hz, DABA1-α-CH),4.39-4.24(3H,m,H-2’+H-4’+Trp4-α-CH),4.24-4.08(3H,m,DABA1-β-CH+ Nal2-α-CH+Leu3-α-CH),3.34-3.28(1H,m,H-5’),3.12-2.96(3H,H-5’+Trp4-β-CH2),2.91- 2.86(1H,m,Nal2-β-CH2), 2.81 (1H, dd, J=13.2,6.7Hz, Nal2- β-CH2),2.60(3H,s,NCH3), 2.58-2.52(1H,m,Nal2-β-CH2), 2.10 (1H, app.dt, J=13.4,6.8Hz, H-3 '), 1.68-1.50 (2H, m, H-3’+Leu3-γ-CH),1.36-1.28(2H,m,Leu3-β-CH2),0.89-0.76(6H,m,2×Leu3-δ-CH3),0.63 (3H, d, J=6.5Hz, DABA1- γ-CH3).13CNMR(125MHz,DMSO-d6, the ratio 1.8:1 of rotational isomer, mainly Rotational isomer): δ 174.3 (C=O), 172.8 (C=O), 169.9 (C=O), 164.6 (C=O), 163.2 (C=O), 157.3 (C=O), 150.4 (C=O), 141.8,136.2,135.9,135.7,132.0,131.7,128.0,127.8, 127.6,127.5,127.4(×2),127.3,125.4,123.4,120.4,118.4,118.0,111.0,110.0,101.4, 92.1,78.8,74.0,55.2,54.1,51.8,51.6,43.4,41.2,40.1,35.2,27.9,27.3,23.7,21.8, 14.5.LRMS[M+H+]882.4.HRMS(ESIm/z)[M+H+]C45H55N9O10Calculated value 882.4144, measured value 882.4140.
(((S) -1- (((2S, 3S) -3- ((S) -2- amino -3- cyclohexyl-N-methyl propionamide) -1- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxygen For butane-2- base) amino)-4- methyl-1-oxo-pentane-2- base) carbamoyl)-L-Trp (10)
According to the condition B of general step 8, at room temperature, HOAt (17mg, 123 μm of ol) and EDCHCl (5.2mg, 25 μ Mol) and in the presence of NMM (3 μ L, 25 μm of ol), depsipeptide S28 (22mg, 25 μm of ol) and amine 18 (6.8mg, 30 μm of ol) is made to exist CH2Cl2: reaction 3 hours in DMF (1:1v/v, 240 μ L) obtain the analog protected completely.According to general step 8, the chemical combination Object is with TFA and containing iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 2.5mL), reversed-phase HPLC (0 to 50%MeCN, 40 minutes；100%H2O, 10 minutes) amorphous white solid is obtained after purification 10 (tfa salts) (10.4mg, two steps, 44%).
IR(ATR):3346,2924,1728,1627cm-1.1HNMR(500MHz,CD3OD, the ratio of rotational isomer 1.6:1, main rotational isomer): δ 8.67 (1H, t, J=6.0Hz), 8.45 (1H, d, J=9.0Hz), 8.16-8.05 (1H, m), 7.62 (1H, d, J=8.1Hz, H-6), 7.58 (1H, dd, J=7.9,4.0Hz, Ar-H), 7.33 (1H, dd, J= 8.1,5.6Hz, Ar-H), 7.14-7.05 (2H, m, Ar-H), 7.03-6.98 (1H, m, Ar-H), 5.74 (1H, d, J=2.7Hz, H-1 '), 5.70 (1H, d, J=8.0Hz, H-5), 4.88-4.78 (1H, m, DABA1- β-CH), 4.67-4.54 (3H, m, DABA1-α-CH+Cha2-α-CH+Trp4-α-CH),4.49-4.38(2H,m,H-2’+H-4’),4.19-4.08(1H,m, Leu3- α-CH), 3.52-3.46 (1H, m, H-5 '), 3.29-3.23 (2H, m, H-5 ' main+Trp4- β-CH2),3.24- 3.14(1H,m,Trp4-β-CH2),2.85(3H,s,NCH3),2.30-2.18(1H,m,H-3’),1.84-1.55(10H,m,4× CH2+H-3’+Leu3-γ-CH),1.52-1.40(3H,CH2+Cha2-γ-CH),1.36-1.26(2H,m,Leu3-β-CH2), 1.24-1.10(3H,m,DABA1-γ-CH3),1.07-0.95(2H,m,CH2),0.94-0.86(6H,m,2×Leu3-δ- CH3).13CNMR(125MHz,CD3OD, the ratio 1.6:1 of rotational isomer, main rotational isomer): δ 176.2 (C=O), 175.6 (C=O), 171.8 (C=O), 171.0 (C=O), 166.3 (C=O), 159.8 (C=O), 152.3 (C=O), 142.2,137.9,128.6,124.4,122.0,119.4,119.2,111.9,110.6,102.2,95.0,80.9,76.1, 54.9,54.0,52.6,50.3,44.8,41.8,39.6,36.0,34.8,34.2,29.0,28.3,26.9,25.6,22.9, 14.6.LRMS[M+H+]838.4.HRMS(ESIm/z)[M+Na+]C46H72N6O11Na calculated value 907.5151, measured value 907.5150。
(((S) -1- (((2S, 3S) -3- ((S) -2- amino -2- cyclohexyl-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxygen For butane-2- base) amino)-4- methyl-1-oxo-pentane-2- base) carbamoyl)-L-Trp (11)
According to the condition A of general step 8, at room temperature, HOAt (14mg, 107 μm of ol) and DIC (5.2 μ L, 33 μm of ol) are deposited Under, by depsipeptide S29 (19mg, 22 μm of ol) and amine 18 (5.8mg, 26 μm of ol) in CH2Cl2: DMF (1:1v/v, 200 μ L) Middle reaction 18 hours obtains protection analog completely.According to universal method 8, the compound is with TFA and contains iPr3SiH's CH2Cl2Mixture handles (1:1v/vTFA:CH2Cl2, 2.5vol.%iPr3SiH, 2.2mL), reversed-phase HPLC (0 to 50%MeCN, 40 minutes；10 minutes, 100%H2O amorphous white solid 11 (tfa salt) (5.4mg, 2 steps, yield 26%)) are obtained after purification.
IR(ATR):3261,2931,2856,1665cm-1.1HNMR(400MHz,CD3The rotational isomeric of OD, 1.6:1 ratio Body, main rotational isomer): δ 10.30 (1H, s, NH), 8.67 (1H, s, NH), 8.12 (1H, s, NH), 7.62 (1H, d, J =8.1Hz, H-6), 7.58 (1H, dd, J=8.0,3.0Hz, Ar-H), 7.39-7.24 (1H, m, Ar-H), 7.18-7.03 (2H, M, Ar-H), 7.04-6.98 (1H, m, Ar-H), 5.74 (1H, d, J=2.7Hz, H-1 '), 5.71 (1H, d, J=8.0Hz, H- 5),4.91-4.78(1H,m,DABA1-β-CH),4.71-4.53(1H,Trp4-α-CH),4.51-4.37(2H,m,H-2’+H- 4 '), 4.30-4.22 (1H, m, DABA1- α-CH), 4.20-4.08 (1H, m, Leu3- α-CH), 4.03 (1H, d, J=4.6Hz, Chg2- α-CH), 3.55-3.45 (1H, m, H-5 '), 3.29-3.25 (2H, m, H-5 ' main+Trp4- β-CH2),3.22- 3.09(1H,m,Trp4-β-CH2),2.86(3H,s,NCH3),2.35-2.16(1H,m,H-2’),1.87-1.54(6H,m,H-2’ +Chg2-β-CH+1.5×Chg2-CH2+Leu3-γ-CH),1.53-1.42(2H,m,Leu3-β-CH2),1.35-1.02(6H, m,DABA1-γ-CH3+1.5×Chg2-CH2, main), 0.91 (6H, m, 2 × Leu3- δ-CH3It is main)13CNMR (100MHz,CD3OD, main rotational isomer): δ 176.1 (C=O), 175.1 (C=O), 171.4 (C=O), 169.7 (C =O), 166.0 (C=O), 159.5 (C=O), 151.9 (C=O), 142.3,137.8,128.6,124.7,122.0,119.6, 119.2,111.9,110.5,109.1,102.3,95.0,80.8,76.2,56.4,55.0,54.4,53.8,52.8,44.6, 43.6,42.0,40.5,35.8,30.2,29.0,28.3,26.7,25.7,23.4,15.2.LRMS[M+H+]824.4.HRMS (ESIm/z)[M+H+]C40H57N9O10Calculated value 824.4301, measured value 824.4294.
(((2S, 3R) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) (methyl) amino) -1- oxo-butanes -2- Base) amino)-3- methyl-1-oxo-pentane-2- base) carbamoyl)-L-Trp (21)
According to the condition B of general step 8, at room temperature, HOAt (15mg, 108 μm of ol) and EDCHCl (5.4mg, 28 μ Mol) and in the presence of NMM (3.0 μ L, 28 μm of ol), make depsipeptide S36 (17mg, 22 μm of ol) and amine 18 (7.4mg, 32 μm of ol) In CH2Cl2: DMF (1:1v/v, 210 μ L) reacts 4 hours, obtains the analog protected completely.According to universal method 8, the chemical combination Object is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/vTFA:CH2Cl2, 2.5vol.%iPr3SiH, 2.2mL) 4 hours.According to general step 8, and reversed-phase HPLC (0 to 50%MeCN, 45 minutes；100%H2O, 10 minutes, flow velocity 9mL/min) After purification, amorphous white solid 21 (such as formates) (7.4mg, 44%, 2 step) is obtained.
IR(ATR):3374,2979,2932,1731,1666cm-1.1HNMR(500MHz,DMSO-d6, rotational isomer Ratio 1:1, rotational isomer 1): δ 10.77 (s, 1H), 8.37 (s, 1H), 8.05 (s, 1H), 7.62-7.47 (2H, m, Ar-H+ ), H-6 7.30 (1H, dd, J=8.1,5.0Hz, Ar-H), 7.13 (1H, s, Ar-H), 7.09 (1H, s, Ar-H), 7.04-6.99 (1H, m, Ar-H), 6.94 (1H, app.td, J=7.2,3.0Hz, Ar-H), 6.40 (1H, d, J=8.6Hz), 6.22 (1H, s), 5.70 (1H, d, J=3.6Hz, H-1 '), 5.60-5.56 (1H, m, H-5), 4.59-4.52 (1H, m, DABA1- α-CH), 4.40- 4.16(3H,m,H-2’+H-4’+Trp4-α-CH),4.04-3.89(2H,m,DABA1-β-CH+Ile3-α-CH),3.67(1H, D, J=16.2Hz, Gly2- α-CH2),3.62-3.54(1H,m,Gly2-α-CH2), 3.41 (1H, J=14.5Hz), 3.28- 3.04(3H,m,H-5’+Trp4-β-CH2), 3.00 (1H, dd, J=14.5,6.3Hz, Trp4- β-CH2),2.70(3H,s, NCH3),2.20-2.07(1H,m,H-3’),1.80-1.56(2H,m,H-3’+Ile3-β-CH),1.47-1.34(1H,m, Ile3-γ-CH2), 1.11 (3H, d, J=6.5Hz, DABA1- γ-CH3),1.08-0.98(1H,m,Ile3-γ-CH2),0.78 (6H,m,2×CH3).13CNMR(125MHz,DMSO-d6, the ratio 1:1 of rotational isomer, rotational isomer 1, have one it is fuzzy 's13C signal): δ 174.8 (C=O), 172.2 (C=O), 169.8 (C=O), 169.3 (C=O), 163.2 (C=O), 150.6 (C=O), 141.1,136.0,127.5,123.4,120.6,118.4,117.8,111.5,110.3,10 1.5,91.9,78.2, 72.7,57.4,55.3,54.5,52.3,43.2,40.4,39.6,36.8,35.2,27.7,24.2,14.9,14.5, 10.9.1HNMR(500MHz,DMSO-d6, the ratio 1:1 of rotational isomer, rotational isomer 2): δ 8.52 (s, 1H), 8.18 (d, J=9.0Hz, 1H), 5.65 (1H, d, J=3.6Hz, H-1 '), 4.82-4.74 (1H, m, DABA1- β-CH), 4.51-4.44 (1H,m,DABA1-α-CH),2.72(3H,s,NCH3).13CNMR(125MHz,DMSO-d6, the ratio 1:1 of rotational isomer, rotation Turn isomers 2): δ 50.0.LRMS [M+H+]742.4.HRMS(ESIm/z)[M+H+]C34H48N9O10Calculated value 742.3518 is surveyed Magnitude 742.3516.
(2S, 6S, 9S, 10S) -2- ((1H- indol-3-yl) methyl) -13- amino -9- ((((2R, 4R, 5R) -5- (2,4- - 1 (2H)-yl of dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) carbamoyl) -6- isopropyl - Four azepine tridecane -1- of 10,11- dimethyl -4,7,12- trioxy- -3,5,8,11- is sour (22)
According to the condition B of general step 8, at room temperature, HOAt (15.7mg, 115 μm of ol), EDCHCl (5.8mg, 30 μ Mol) and in the presence of NMM (3.3 μ L, 30 μm of ol), make depsipeptide S37 (17.8mg, 23.4 μm of ol) and amine 18 (10.5mg, 46 μ Mol) in CH2Cl2: reaction 4 hours in DMF (1:1v/v, 240 μ L) obtain the analog protected completely.According to universal method 8, The compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/vTFA:CH2Cl2, 2.5vol.%iPr3SiH, 1.2mL) 16 hours, it is indissociable diastereomeric different that reversed-phase HPLC purifies (0 to 50%MeCN, 40 minutes, 0.1vol% formic acid) The 9:1 mixture of structure body, (8.5mg, 62%, are based on diastereoisomer formic acid in HPLC to fluffy white solid 22 after purification The yield that salt calculates).Therefore have submitted inseparable diastereoisomer 9:1 mixture for characterize and bioassay.
IR(ATR):3316,2925,2872,1672,1558cm-1.1HNMR(500MHz,DMSO-d6, rotational isomer Ratio 1.4:1, rotational isomer 1): δ 7.52 (2H, m, H-6+Ar-H), 7.30 (1H, m, Ar-H), 7.09 (1H, m, Ar-H), 7.02(1H,m,Ar-H),6.92(1H,m,Ar-H),5.64(1H,m,H-1’),5.59(1H,m,H-5),4.57(1H,m, DABA1-α-CH),4.30(2H,m,H-2’+H-4’+Val3-α-CH+Trp4-α-CH),3.98(1H,m,DABA1-β-CH), 3.89(2H,m,Gly2-α-CH2),3.22-3.00(4H,m,Trp4-β-CH2+2xH-5’),2.73(3H,s,NCH3),2.14– 2.11(1H,m,H-3’),1.63-1.60(1H,m,H-3’),1.11(3H,m,DABA1-γ-CH3),1.04(1H,m,Val3- β-CH),0.79(6H,m,2xVal-γ-CH3).13CNMR(126MHz,DMSO-d6, the ratio 1.4:1 of rotational isomer, rotation Isomers 1): δ 173.7 (C=O), 172.4 (C=O), 170.1 (C=O), 167.2 (C=O), 163.3 (C=O), 157.9 (C =O), 151.0 (C=O), 141.4,136.5,128.0,123.6,120.9,118.9,111.7,110.7,101.4,91 .9, 78.6,74.1,58.4,55.0,54.6,52.9,43.7,40.0,35.9,31.4,29.1,27.6,19.8,18.1,15.1, 14.5.1HNMR(500MHz,DMSO-d6, the ratio 1.4:1 of rotational isomer, rotational isomer 2): δ 5.67 (1H, m, H-1 ') .13CNMR(126MHz,DMSO-d6, the ratio 1.4:1 of rotational isomer, rotational isomer 2): δ 92.3.LRMS [M+H+] 728.4.HRMS(ESIm/z)[M+H+]C33H45N9O10Calculated value 728.3362, measured value: 728.3359.
(((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- base) ammonia Base) -1- oxo heptane -2- base) carbamoyl)-L-Trp (23)
According to the condition B of general step 8, at room temperature, HOAt (25mg, 185 μm of ol) and EDCHCl (9.2mg, 48 μ Mol) and in the presence of NMM (5.0 μ L, 48 μm of ol), depsipeptide S38 (30mg, 37 μm of ol) and amine 18 (20mg, 92 μm of ol) is made to exist CH2Cl2: reaction 4 hours in DMF (1:1v/v, 370 μ L) obtain the analog protected completely.According to universal method 8, the chemical combination Object is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 3.7mL) 4 hours, and reversed-phase HPLC (0 to 50%MeCN, 40 minutes；H2O%, 10 minutes) after purification, it is solid to obtain amorphous white Body 23 (as formates) (8.1mg, 2 steps, yield 27%).
IR(ATR):3356,2977,2932,1731,1638cm-1.1HNMR(400MHz,DMSO-d6, rotational isomer Ratio 1:1, rotational isomer 1): δ 10.79 (1H, s), 8.64 (s, 1H), 8.37 (s, 1H), 8.23 (s, 1H), 8.16 (s, 1H),7.54(2H,m,H-6+Ar-H),7.33-7.28(1H,m,Ar-H),7.13(1H,s,Ar-H),6.97-6.91(1H,m, ), Ar-H 6.45 (1H, d, J=7.8Hz), 6.15 (1H, s), 5.71 (1H, d, J=3.3Hz, H-1 '), 5.58 (1H, app.dd, J=8.0,2.5Hz, H-5), 4.55 (1H, app.t, J=9.4Hz, DABA1- α-CH), 4.24-4.17 (3H, m, H-2 '+H-4 ' +Trp4-α-CH),4.09-4.00(1H,m,Aha3-α-CH),3.99-3.88(1H,m,DABA1-β-CH),3.32-3.00 (3H,m,H-5’+Trp4-β-CH2),3.70-3.61(1H,m,Gly2-α-CH2), 3.46 (1H, d, J=14.9Hz, Gly2- α- CH2), 2.99 (1H, dd, J=14.6,6.5Hz, Trp4- β-CH2),2.72(3H,s,NCH3),2.71(3H,s,NCH3), 2.15-2.05(1H,m,H-3’),1.71-1.63(1H,m,H-3’),1.56-1.37(2H,m,Aha3-β-CH2),1.30- 1.14 (6H, m), 1.10 (3H, d, J=6.5Hz, DABA1- γ-CH3),0.86-0.77(3H,m).13CNMR(100MHz, DMSO-d6, the ratio 1:1 of rotational isomer, rotational isomer 1): δ 175.2 (C=O), 173.9 (C=O), 170.1 (C= ), O 169.8 (C=O), 168.8 (C=O), 163.3 (C=O), 150.9 (C=O), 141.5,136.4,128.0,124.2, 123.8,120.7,117.8,111.6,110.6,101.2,92.0,78.4,73.8,54.3,53.4,50.7,42.9,40.2, 34.7,31.2,28.6,28.4,24.9,21.8,15.3,13.1.1HNMR(400MHz,DMSO-d6, the ratio of rotational isomer 1:1, rotational isomer 2): δ 8.06 (s, 1H), 7.06-6.99 (1H, m, Ar-H), 7.10 (1H, app.d, J=2.3Hz, Ar- H), 5.65 (1H, d, J=3.4Hz, H-1 '), 4.82-4.73 (1H, m, DABA1- β-CH), 4.49-4.42 (1H, m, DABA1- α-CH), 4.41-4.34 (1H, m, H-4 '), 2.15-2.05 (1H, m, H-3 '), H-3 ' is secondary), 1.77 (1H, app.d, J= 13.0Hz,H-3’).13CNMR
(100MHz,DMSO-d6, the ratio 1:1 of rotational isomer, rotational isomer 2): δ 27.0.LRMS [M+H+] 756.4.HRMS(ESIm/z)[M+H+]C35H50N9O10Calculated value 756.3675, measured value 756.3674.
(((S) -2- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1-- oxo-butanes -2- base) Amino) -1- cyclohexyl -2- oxoethyl) carbamoyl)-L-Trp (24)
According to the condition B of general step 8, at room temperature, HOAt (20mg, 150 μm of ol) and EDCHCl (7.4mg, 39 μ Mol) and in the presence of NMM (4.0 μ L, 39 μm of ol), depsipeptide S39 (24mg, 30 μm of ol) and amine 18 (14mg, 60 μm of ol) is made to exist CH2Cl2: reaction 4 hours in DMF (1:1v/v, 300 μ L) obtain the analog protected completely.According to universal method 8, the chemical combination Object is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/vTFA:CH2Cl2, 2.5vol.%iPr3SiH, 3.0mL) 4 hours, reversed-phase HPLC (0 to 50%MeCN, 40 minutes；10 minutes, 100%H2O) after purification, amorphous white solid is obtained 24 (tfa salts) (13.7mg, 2 steps, yield 60%).
IR(ATR):3323,2979,2932,1626cm-1.1HNMR(400MHz,CD3OD, the ratio of rotational isomer 1.6:1, main rotational isomer): δ 8.78 (1H, t, J=5.9Hz), 8.37 (1H, d, J=8.9Hz), 8.22 (1H, d, J =8.5Hz), 8.15 (1H, t, J=5.8Hz), 7.62 (1H, d, J=8.1Hz, H-6), 7.58-7.54 (1H, m, Ar-H), 7.38-7.26(1H,m,Ar-H),7.16-7.04(2H,m,Ar-H),7.04-6.95(1H,m,Ar-H),5.77-5.70(2H, m,H-1’+H-5),4.62-4.56(2H,m,DABA1-α-CH+Trp4-α-CH),4.50-4.42(2H,m,H-2’),4.39- 4.31 (1H, m, H-4 '), 4.16 (1H, d, J=16.0Hz, Gly2- α-CH2),4.03-3.88(3H,m,DABA1-β-CH+ Gly2-α-CH2+Chg3-α-CH),3.54-3.46(1H,m,H-5’),3.29-3.13(3H,m,H-5’+Trp4-β-CH2), 2.87(3H,s,NCH3),2.78(3H,s,NCH3), 2.21 (1H, ddd, J=13.6,7.6,6.1Hz, H-3 '), 1.84-1.48 (6H,m,H-3’+2.5×CH2+Chg3-β-CH),1.25-0.95(8H,m,DABA1-γ-CH3+2.5×CH2).13CNMR (100MHz,CD3OD, the ratio 1.6:1 of rotational isomer, main rotational isomer): δ 176.2 (C=O), 175.0 (C= ), O 171.3 (C=O), 167.6 (C=O), 167.2 (C=O), 160.0 (C=O), 152.2 (C=O), 142.4,137.9, 128.8,124.2,122.1,119.6,119.4,111.8,110.4,102.0,94.7,81.2,75.8,60.0,56.6, 54.6,54.1,45.0,41.5,40.8,35.6,30.6,29.6,29.0,27.8,26.8,14.9.LRMS[M+H+] 768.3.HRMS(ESIm/z)[M+H+]C36H50N9O10Calculated value 768.3875, measured value 768.3867.
(((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- base) ammonia Base) -3- cyclohexyl -1- oxopropan -2- base) carbamoyl)-L-Trp (25)
According to the condition B of general step 8, at room temperature, HOAt (14mg, 103 μm of ol) and EDCHCl (5.1mg, 27 μ Mol) and in the presence of NMM (3.0 μ L, 27 μm of ol), make depsipeptide S40 (17mg, 20 μm of ol) and amine 18 (9.5mg, 42 μm of ol) In CH2Cl2: reaction 4 hours in DMF (1:1v/v, 200 μ L) obtain the analog protected completely.According to universal method 8, the change It closes object and with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 4mL) 4 hours, reversed-phase HPLC (0 to 50%MeCN, 45 minutes；10 minutes, 100%H2O amorphous white) is obtained after purification Solid 25 (6.0mg, 2 steps, yield 35%).
IR(ATR):3368,2926,1647,1549cm-1.1H NMR(600MHz,CD3OD, the ratio of rotational isomer 1.5:1, main rotational isomer): δ 7.62 (1H, d, J=8.1Hz, H-6), 7.60-7.54 (1H, m, Ar-H), 7.33 (1H, dd, J=9.1,8.2Hz, Ar-H), 7.11 (1H, s, Ar-H), 7.10-7.05 (1H, m, Ar-H), 7.04-6.98 (1H, M, Ar-H), 5.73 (1H, d, J=2.6Hz, H-1 '), 5.71 (1H, d, J=8.0Hz, H-5), 4.65 (1H, d, J=7.6Hz, DABA1-α-CH),4.60-4.52(2H,m,H-4’+Trp4-α-CH),4.49-4.44(1H,m,H-2’),4.21-4.12(2H, m,Gly2-CH2+ Cha3- α-CH), 3.99 (1H, dq, J=10.1,6.6Hz, DABA1- β-CH), 3.91 (1H, d, J= 16.0Hz,Gly2-CH2),3.55-3.49(1H,dd,m,H-5’),3.29-3.16(2H,m,Trp4-β-CH2),2.88(3H,s, NCH3), 2.22 (1H, ddd, J=13.8,7.7,6.2Hz, H-3 '), 1.84-1.60 (5H, m, 2 × Cha3-CH2+H-3’), 1.49-1.41(2H,m,Cha3-β-CH2),1.36-1.31(1H,m,Cha3-γ-CH),1.26-1.09(7H,m,2×Cha3- CH2+DABA1-γ-CH3),1.03-0.79(2H,m,Cha3-CH2).13C NMR(150MHz,CD3The ratio of OD rotational isomer 1.5:1, main rotational isomer): δ 176.3 (C=O), 175.9 (C=O), 171.9 (C=O), 167.4 (C=O), 167.1 (C=O), 160.2 (C=O), 152.5 (C=O), 142.6,138.1,129.1,124.5,122.0,119.3, 119.0,112.0,110.4,102.3,95.1,80.6,76.2,56.6,54.8,54.4,53.0,44.8,40.9,40.6, 36.2,34.9,34.4,28.5,28.4,27.5,26.8,13.9.LRMS[M+H+]782.4.HRMS(ESI m/z)[M+H+] C37H52N9O10Calculated value 782.3831, measured value 782.3824.
(((2S, 3S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- Base) amino) -3- hydroxyl -1- oxo-butanes -2- base) carbamoyl)-L-Trp (26)
According to general step 8, at room temperature, HOAt (15.8mg, 116.0 μm of ol), EDCHCl (5.8mg, 30.1 μ Mol) and in the presence of NMM (3.3 μ L, 30.1 μm of ol), make depsipeptide S41 (18.0mg, 23.2 μm of ol) and amine 18 (10.5mg, 46.4 μm of ol) in CH2Cl2: DMF (1:1v/v, 240 μ L) reaction obtains the analog protected completely.It, should according to universal method 8 Compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 1.2mL) 16 hours, fluffy white solid 26 (formates) was obtained after reversed-phase HPLC purifies (0 to 30%MeCN, 40 minutes) (8.3mg, two steps, 46%).
IR(ATR):3293,2922,1677,1594,1436cm-1.1H NMR(500MHz,DMSO-d6, 1.3:1 ratio Rotational isomer, rotational isomer 1): δ 7.53 (2H, m, H-6+Ar-H), 7.30 (1H, m, Ar-H), 7.12 (1H, m, Ar-H), 7.03 (1H, m, Ar-H), 6.94 (1H, m, Ar-H), 6.55 (1H, br s, Thr3- γ-OH), 5.64 (1H, d, J=3.1Hz, H-1 '), 5.57 (1H, d, J=6.4Hz, H-5), 4.60 (1H, m, DABA1- α-CH), 4.30 (1H, m, Trp4- α-CH), 4.27–4.23(2H,m,H-2’+H-4’),4.00–3.94(3H,m,Thr3-α-CH+Thr3-β-CH+DABA1-β-CH), 3.36–3.00(6H,m,Gly2-α-CH2+2x H-5’+Trp4-β-CH2),2.49(3H,s,NCH3),2.14–2.11(1H,m, H-3 '), 1.71-1.62 (1H, m, H-3 '), 1.15 (3H, d, J=4.7Hz, Thr3- γ-CH3), 0.99 (3H, d, J= 5.5Hz,DABA1-γ-CH3).13C NMR(126MHz,DMSO-d6, the ratio 1.3:1 of rotational isomer, rotational isomer 1): δ 179.3 (C=O), 173.7 (C=O), 171.8 (C=O), 170.0 (C=O), 164.5 (C=O), 163.6 (C=O), 150.8 (C=O), 141.7,136.4,127.9,124.3,121.2,119.2,118.5,111.7,110.0,10 2.1,92.6, 79.0,78.9,74.1,66.9,59.4,55.3,55.2,44.1,40.3,35.8,29.2,27.6,20.1,15.1.1H NMR (500MHz,DMSO-d6；The ratio 1.3:1 of rotational isomer, rotational isomer 1): δ 5.68 (1H, m, H-1 '), 5.58 (1H, m,H-5).LRMS[M+H+]C32H44N9O11Calculated value 730.3154, measured value 730.3150.
(2S, 6S, 9S, 10S) -2- ((1H- indol-3-yl) methyl) -13- amino -6- (carboxymethyl) -9- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) carbamyl Base) four azepine tridecanoic acid (27) of -10,11- dimethyl -4,7,12- trioxy- -3,5,8,11-
According to 8 condition B of general step, at room temperature, HOAt (14.7mg, 107.8 μm of ol), EDCHCl (5.4mg, 28.0 μm of ol) and NMM (3.1 μ L, 28.0 μm of ol), make depsipeptide S42 (19.2mg, 21.6 μm of ol) and amine 18 (12.2mg, 53.9 μm of ol) in CH2Cl2: 4 hours condition B of reaction obtain the analog protected completely in DMF (1:1v/v, 220 μ L).According to logical With method 8, the compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 1.2mL) 16 hours, after reversed-phase HPLC (0 to 50%MeCN, 40 minutes), obtain white solid 27 (formates) (10.0mg, two steps, 62%).
IR(ATR):3304,2952,2844,1643,1450,1411cm-1.1H NMR(500MHz,DMSO-d6, rotate different The ratio 1.1:1 of structure body, rotational isomer 1): δ 7.52 (2H, m, H-6+Ar-H), 7.31 (1H, m, Ar-H), 7.12 (1H, s, Ar-H),6.99(1H,m,Ar-H),6.93(1H,m,Ar-H),5.65(1H,m,H-5),5.57(1H,m,H-1’),4.41(1H, m,Asp3-α-CH),4.33-4.28(4H,m,H-2’+H-4’+DABA1-α-CH+Trp4-αCH),3.94–3.55(2H,m, Gly2-α-CH2),3.27(1H,m,DABA1-β-CH),3.12(1H,m,H-5’),2.99(1H,m,H-5’),2.71(3H,m, NMe),2.44(4H,m,Asp3-β-CH2+Trp4-β-CH2),2.15–2.11(1H,m,H-3’),1.68–1.65(1H,m,H- 3’),1.08(3H,m,DABA1-γ-CH3).1H NMR(500MHz,DMSO-d6, the ratio 1.1:1 of rotational isomer, rotate it is different Structure body 2): δ 5.59 (1H, m, H-1 '), 5.66 (1H, m, H-5), 2.65 (3H, m, NMe), 1.75-1.68 (1H, m, H-3 ')13C NMR:(126MHz,DMSO-d6): δ 179.5 (C=O), 172.1 (C=O), 167.8 (C=O), 166.8 (C=O), 166.1 (C =O), 163.8 (C=O), 163.7 (C=O), 150.9 (C=O), 141.5,136.3,128.0,124.0,123.9,121.0, 118.9,111.6,110.7,110.9,101.9,92.5,78.8,74.4,55.6,55.0,51.2,43.9,40.2,40.0, 35.9,29.4,28.5,27.8,15.0. rotational isomer 2: undistinguishable .LRMS [M+H+]744.4.HRMS(ESI m/z)[M +H]+C32H42N9O12Calculated value 744.2947, measured value 744.2943.
(((S) -6- amino -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes - 2- yl) amino) -1- oxohexane -2- base) carbamoyl)-L-Trp (28)
According to the condition B of general step 8, at room temperature, HOAt (10mg, 74.0 μm of ol), EDCHCl (3.7mg, 19.2 μm ol) and NMM (2.1 μ L, 19.2 μm of ol) in the presence of, make depsipeptide S43 (13.4mg, 14.8 μm of ol) and amine 18 (11mg, 48.7 μm of ol) in CH2Cl2: reaction 4 hours in DMF (1:1v/v, 180 μ L) obtain corresponding shielded analog.Then it presses According to universal method 8, the compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 1.2mL) 16 hours, it is solid that fluffy white is obtained after reversed-phase HPLC (0 to 30%MeCN, 40 minutes) Body 28 (tfa salt) (7.2mg, two steps, 62%).
IR(ATR):3285,3083,2941,1672,1555cm-1.1H NMR(500MHz,DMSO-d6, rotational isomer Ratio 2.2:1, main rotational isomer): δ 7.55-7.49 (2H, m, H-6+Ar-H), 7.32 (1H, d, J=8.0Hz, Ar-H),7.11(1H,s,Ar-H),7.04(1H,m,Ar-H),6.95(1H,m,Ar-H),5.68–5.57(2H,m,H-5+H- 1’),4.79–4.73(1H,m,Lys3-α-CH),4.36-4.32(3H,m,H-2’+DABA1-α-CH+H-4’),4.11–4.09 (1H,m,Trp4-α-CH),3.89–3.86(1H,m,DABA1-β-CH),3.86–3.65(2H,m,Gly2-α-CH2),3.25– 3.24(1H,m,H-5’),3.19–2.98(3H,m,H-5’+2x Trp4-β-CH2),2.74(5H,m,NMe+Lys3-ε-CH2), 2.21–2.08(1H,m,H-3’),1.51-1.48(3H,m,H-3’+Lys3-δ-CH2),1.27–1.24(2H,m,Lys3-γ- CH2), 1.11 (3H, d, J=6.4Hz, DABA1- γ-CH3), 1.02 (2H, d, J=6.7Hz, Lys3- β-CH2).13C NMR (126MHz,DMSO-d6, the ratio 2.2:1 of rotational isomer, main rotational isomer): δ 173.7 (C=O), 172.0 (C =O), 169.8 (C=O), 169.0 (C=O), 165.6 (C=O), 162.8 (C=O), 150.0 (C=O), 141.7,135.6, 126.9,124.0,121.2,118.7,111.7,109.0,79.0,74.3,55.4,55.1,54.8,53.7,53.1,44.0, 40.1,39.1,36.0,32.2,29.0,28.1,27.7,27.0,22.7,15.0,14.3.LRMS[M+H+]757.4.HRMS (ESI m/z)[M+H+]C34H49N10O10Calculated value 757.3627, measured value 757.3620.
(((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- iso-butane -2- base) ammonia Base) -1- oxo -3- (4- (trifluoromethyl) phenyl) propane -2- base) carbamoyl)-L-Trp (30)
According to the condition B of general step 8, at room temperature, HOAt (22mg, 161 μm of ol) and EDCHCl (8.0mg, 42 μ Mol) and in the presence of NMM (4.4 μ L, 42 μm of ol), depsipeptide S45 (28mg, 32 μm of ol) and amine 18 (15mg, 64 μm of ol) is made to exist CH2Cl2: reaction 4 hours in DMF (1:1v/v, 320 μ L) obtain the analog protected completely.According to universal method 8, the chemical combination Object is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 3.2mL) 4 hours, reversed-phase HPLC (0 to 50%MeCN, 45 minutes；10 minutes, 100%H2O it) obtains after purification amorphous white Color solid 30 (formates) (5.6mg, 2 steps, 20%).
IR(ATR):3331,2948,2837,1668cm-1.1H NMR(500MHz,DMSO-d6, the ratio of rotational isomer 1:1, rotational isomer 1): δ 10.73 (1H, s), 8.64 (1H, s), 8.45-8.29 (1H, m), 8.21 (1H, s), 7.66-7.42 (4H,H-6+Ar-H),7.37-7.29(2H,m,Ar-H),7.30-7.16(1H,m,Ar-H),7.05(1H,s,Ar-H),6.97 (1H, app.t, J=7.5Hz, Ar-H), 6.88 (1H, app.t, J=7.5Hz, Ar-H), 6.52 (1H, s), 6.22 (1H, s), 5.76-5.59 (1H, m, H-1 '), 5.55 (1H, d, J=8.0, H-5), 4.54 (1H, app.t, J=9.1Hz, DABA1- α- CH),4.43-4.20(3H,m,H-2’+H-4’+p-CF3-Pal3-α-CH),4.19-4.08(1H,m,Trp4-α-CH),3.94- 3.82(1H,m,DABA1-β-CH),3.79-3.86(1H,m,Gly2-α-CH2),3.60-3.34(1H,m,Gly2-α-CH2), 3.61-3.39(m,1H),3.34-2.89(5H,m,H-5’+p-CF3-Pal3-β-CH2+Trp4-β-CH2),2.88-2.78(1H, m,p-CF3-Pal3-β-CH2),2.70(3H,s,NCH3),2.19-2.04(1H,m,H-3’),1.73-1.53(1H,m,H-3’), 1.03 (3H, d, J=6.0Hz, DABA1- γ-CH3).13C NMR(125MHz,DMSO-d6, the ratio 1:1 of rotational isomer, rotation Turn isomers 1): δ 175.1 (C=O), 173.0 (C=O), 170.9 (C=O), 169.0 (C=O), 167.0 (C=O), 163.1 (C=O), 150.4 (C=O), 142.1,140.6,135.3,129.4,127.4,126.6,123.9,123.6,119.7, 118.0,117.4,117.1,110.4,110.3,100.9,91.3,78.6,73.4,54.4,54.1,53.5,52.1,42.8, 39.3,36.8,34.6,27.2,26.5,13.8.1H NMR(500MHz,DMSO-d6, the ratio 1:1 of rotational isomer, rotate it is different Structure body 2): δ 4.77-4.70 (1H, m, DABA1- β-CH), 4.46 (1H, app.t, J=8.7Hz, DABA1- α-CH), 2.68 (3H,s,NCH3).13C NMR(125MHz,DMSO-d6, the ratio 1:1 of rotational isomer, rotational isomer 2): δ 53.5, 49.6,28.0,13.2.LRMS[M+H+]844.3.HRMS(ESI m/z)[M+H+]C38H45F3N9O10Calculated value 844.3260 is surveyed Magnitude 844.3234.
(2S, 6S, 9S, 10S) -2- ((1H- indol-3-yl) methyl) -13- amino -9- ((((2R, 4R, 5R) -5- (2,4- - 1 (2H)-yl of dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) carbamoyl) -6- (4- hydroxyl Benzyl) four azepine tridecane -1- of -10,11- dimethyl -4,7,12- trioxy- -3,5,8,11- are sour (31)
According to general step 8, at room temperature, HOAt (22.5mg, 165.0 μm of ol), EDCHCl (8.2mg, 42.9 μ Mol) and in the presence of NMM (4.7 μ L, 42.9 μm of ol), make depsipeptide S46 (29.5mg, 33.0 μm of ol) and amine 18 (18.7mg, 82.4 μm of ol) in CH2Cl2: reaction 4 hours in DMF (1:1v/v, 340 μ L) obtain shielded analog completely.Then according to Universal method 8, the compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 1.2mL) 16 hours, it is obtained after reversed-phase HPLC purifies (0 to 50%MeCN, 40 minutes) fluffy White solid 31 (formates) (15.7mg, 50%, two step).
IR(ATR):3325,2928,1676,1541,1498cm-1.1H NMR(400MHz,DMSO-d6, rotational isomer Ratio 1.5:1, main rotational isomer): δ 7.53 (2H, m, H-6+Ar-H), 7.30 (1H, d, J=8.0Hz Ar-H), 7.10 (1H, app.dd, J=6.2Hz, Ar-H), 7.03 (1H, m, Ar-H), 6.95 (3H, m, 3xAr-H), 6.61 (2H, m, 2x Ar-H), 5.67 (1H, d, J=3.2Hz, H-1 '), 5.59 (1H, d, J=2.3Hz, H-5), 4.55 (1H, m, DABA1- α-CH), 4.32-4.24(4H,m,Trp4-α-CH+Tyr3-α-CH,H-2’+H-4’),3.91(1H,m,DABA1-β-CH),3.33–2.96 (6H,m,Trp4-β-CH2+Tyr3-β-CH2+2x H-5’),2.82-2.60(5H,m,Gly2-α-CH2+NCH3),2.14–2.11 (1H,m,H-3’),1.70-1.62(1H,m,H-3’),1.10(3H,d,J6.5Hz,DABA1-γ-CH3).13C NMR (101MHz,DMSO-d6, the ratio 1.5:1 of rotational isomer, main rotational isomer): δ 175.0 (C=O), 172.3 (C =O), 169.1 (C=O), 168.3 (C=O), 163.6 (C=O), 159.9,156.2 (C=O), 156.0 (C=O), 141.3, 135.6,130.4,127.3,127.1,124.0,121.1,118.8,118.5,116.2,115.2,109.9,108.8,92.3, 79.0,74.4,55.3,54.2,50.8,43.9,40.6,35.8,29.3,28.3,27.6,15.0.LRMS[M+H+] 792.5.HRMS(ESI m/z)[M+H+] calculated value C37H46N9O11792.3311 measured value 792.3304.
(((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- iso-butane -2- base) ammonia Base) -1- oxo -3- (pyridin-3-yl) propyl- 2- yl) carbamoyl)-L-Trp (32)
According to the condition B of general step 8, at room temperature, HOAt (12mg, 86 μm of ol) and EDC.HCl (4.1mg, 22 μ Mol) and in the presence of NMM (2.3 μ L, 22 μm of ol), depsipeptide S47 (14mg, 17 μm of ol) and amine 18 (10mg, 44 μm of ol) is made to exist CH2Cl2: sustained response 3.5 hours in DMF (1:1v/v, 170 μ L), obtain the analog protected completely.According to universal method 8, The compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.% iPr3SiH, 1.7mL) 4 hours, in reversed-phase HPLC 0 to 50%MeCN, 40 minutes；10 minutes, 100%H2O nothing) is obtained after purification Amorphous white solid 32 (as tfa salt) (8.2mg, 2 step yields 54%).
IR(ATR):3367,3063,1666cm-1.1H NMR(400MHz,CD3OD, the ratio 1.4:1 of rotational isomer, Rotational isomer 1): δ 8.74 (1H, t, J=6.0Hz), 8.60-8.54 (1H, m, Ar-H), 8.41 (1H, d, J=8.8Hz), 8.25-8.16 (1H, m, Ar-H), 7.77-7.67 (1H, m, Ar-H), 7.63 (1H, d, J=8.1Hz, H-6), 7.57-7.47 (1H,m,Ar-H),7.35-7.30(1H,m,Ar-H),7.11-7.04(2H,m,Ar-H),7.03-6.97(1H,m,Ar-H), 5.76-5.74 (1H, m, H-1 '), 5.71 (1H, d, J=8.0Hz, H-5), 4.74-4.62 (1H, m, DABA1- α-CH), 4.61- 4.43(4H,m,H-2’+DABA1-α-CH+3-Pal3-α-CH+Trp4-α-CH),4.41-4.32(1H,m,H-4’),4.14 (1H, d, J=16.0Hz, Gly2- α-CH2),4.01-3.88(2H,m,DABA1-β-CH+Gly2-α-CH2),3.55-3.46 (1H,m,H-5’),3.30-3.10(4H,m,H-5’,3-Pal3-β-CH2+Trp4-β-CH2),3.07-2.97(1H,m,3- Pal3-β-CH2It is main), 2.88 (3H, s, NCH3),2.28-2.18(1H,m,H-3’),1.84-1.69(1H,m,H-3’), 1.25-1.17(3H,m,DABA1-γ-CH3).13C NMR(100MHz,CD3OD, the ratio 1.4:1 of rotational isomer rotate different Structure body 1): δ 176.3 (C=O), 172.8 (C=O), 171.4 (C=O), 167.3 (C=O), 166.8 (C=O), 165.9 (C= ), O 152.2 (C=O), 145.8,145.4,142.8,142.2,137.9 (× 2), 128.8,126.6,124.3,122.1, 119.6,119.2,111.8,110.5,102.2,94.8,81.0,76.1,56.8,55.0,54.8,54.2,44.9,41.0, 36.1,36.0,28.6,27.8,14.8.1H NMR(400MHz,CD3OD, the ratio 1.4:1 of rotational isomer, rotational isomer 2): δ 8.33 (1H, d, J=8.2Hz), 8.15-8.10 (1H, m, Ar-H), 7.57-7.47 (1H, m, Ar-H, H-6), 5.67 (1H, d, J=8.1Hz, H-5), 4.74-4.62 (1H, m, DABA1- β-CH), 4.61-4.43 (1H, m, H-4 '), 3.83 (1H, D, J=16.2Hz, Gly2- α-CH2), 3.75 (1H, d, J=16.2Hz, Gly2- α-CH2),3.46-3.35(1H,m,H-5’), 3.30-3.10(4H,m,H-5’,3-Pal3-β-CH2+Trp4-β-CH2),3.07-2.97(1H,m,3-Pal3-β-CH2),2.81 (3H,s,NCH3), 2.35 (1H, app.dt, J=13.6,7.0Hz, H-3 '), and 1.84-1.69 (1H, m, H-3 '), 1.25-1.17 (3H,m,DABA1-γ-CH3).13C NMR(100MHz,CD3OD, the ratio 1.4:1 of rotational isomer, rotational isomer 2): δ 145.2,102.5,80.4,75.9,56.4,53.3,44.6,41.2,36.4,30.4,13.6.LRMS[M+H+]777.4.HRMS (ESI m/z)[M+H+] calculated value C36H45N10O10Calculated value 777.3314, measured value 777.3311.
Bis- ((1H- indol-3-yl) methyl) -13- amino -9- ((((2R, 4R, the 5R) -5- of (2S, 6S, 9S, 10S) -2,6- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) carbamoyl) -10, Four azepine tridecane -1- of 11- dimethyl -4,7,12- trioxy- -3,5,8,11- is sour (33)
According to general step 8, HOAt (25.5mg, 187.0 μm of ol), EDC.HCl (9.3mg, 48.6 μm of ol) and NMM (5.4 μ L, 48.6 μm of ol) in the presence of, make depsipeptide S48 (36.0mg, 37.4 μm of ol.) exist with amine 18 (21.3mg, 93.5 μm of ol) CH2Cl2: reaction 4 hours in DMF (1:1v/v, 380 μ L) obtain the analog protected completely.It then, should according to universal method 8 Compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 1.2mL) 16 hours, after reversed-phase HPLC (0 to 50%MeCN, 40 minutes), obtain fluffy white solid 33 (formates) (9.1mg, two steps, 32%).
IR(ATR):3327,2964,2928,1680,1554cm-1.1H NMR(400MHz,DMSO-d6, rotational isomer Ratio 1.1:1, rotational isomer 1): δ 7.54 (2H, m, H-6+2x Ar-H), 7.30 (2H, d, J=8.0Hz, 2x Ar-H), 7.11-6.92 (6H, m, 6x Ar-H), 7.02 (1H, m, Ar-H), 6.92 (1H, m, Ar-H), 5.67 (1H, d, J=3.4Hz, H- 1’),5.59(1H,app.d,H-5),4.56(1H,m,DABA1-α-CH),4.48-4.25(4H,m,H-2’+H-4’+Trp3-α- CH+Trp4-α-CH),3.88(1H,m,DABA1-β-CH),3.71(2H,m,Gly2-α-CH2),3.34-2.88(6H,m, Trp3-β-CH2+Trp4-β-CH2+2x H-5’),2.73(3H,s,NCH3),2.14-2.11(1H,m,H-3’),1.76-1.65 (1H,m,H-3’),1.11(3H,m,DABA1-γ-CH3).13C NMR(101MHz,DMSO-d6, the ratio of rotational isomer 1.1:1, rotational isomer 1): δ 173.7 (C=O), 169.0 (C=O), 168.0 (C=O), 167.3 (C=O), 163.7 (C= ), O 163.6 (C=O), 150.8 (C=O), 141.2,136.4,136.3,127.9,127.8,124.3,123.7,121.9, 120.8,119.7,119.5,118.6,118.5,112.2,111.5,111.4,110.2,101.8,92.4,78.8,74.3, 55.2,54.5,51.0,44.0,40.4,39.4,35.8,29.3,28.3,27.2,14.9.1H NMR(400MHz,DMSO-d6, The ratio 1.1:1 of rotational isomer, rotational isomer 2): δ 5.72 (1H, m, H-1 ')13C NMR(101MHz,DMSO-d6, rotation Turn the ratio 1.1:1 of isomers, rotational isomer 2): 93.4,102.8.LRMS [M+H+]815.4.HRMS(ESI m/z)[M+H+] calculated value C39H47N10O10815.3471 measured value 815.3465.
(2S, 6S, 9S, 10S) -2- ((1H- indol-3-yl) methyl) -13- amino -9- ((((2R, 4R, 5R) -5- (2,4- - 1 (2H)-yl of dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) carbamoyl) -10,11- diformazan Three oxygen -6- phenyl -3,5,8,11- of base -4,7,12-, four azepine tridecane -1- is sour (34)
By depsipeptide S49 (25.3mg, 31.3 μm of ol) and amine 18 (17.8mg, 78.2 μm of ol) in CH2Cl2Middle reaction: root According to general step 8, at HOAt (21.3mg, 156.5 μm of ol), EDC.HCl (7.8mg, 40.7 μm of ol) and NMM (4.5 μ L, 40.7 μ Mol in the presence of), the analog protected completely is obtained within DMF (1:1v/v, 320 μ L) 4 hours.Then it according to general step 8, uses CH2Cl2In TFA and i-Pr3The mixture of SiH handles the compound (1:1v/v TFA:CH2Cl2, 2.5 volume %i- Pr3SiH, 1.2mL) 16 hours, 34 (formates) are obtained after reversed-phase HPLC purifies (0 to 50%MeCN, 40 minutes), are fluffy White solid (12.0mg, two steps 47%).
IR(ATR):3407,2923,2853,1679,1437cm-1.1H NMR(400MHz,DMSO-d6, rotational isomer Ratio 1.3:1, rotational isomer 1): δ 7.52 (2H, m, H-6+Ar-H), 7.35-7.29 (6H, m, 6x Ar-H), 7.11- 6.93(3H,m,3x Ar-H),5.64–5.59(2H,m,H-1’+H-5),4.81(1H,m,Phg3-α-CH),4.34(1H,m, DABA1-α-CH),4.32–4.25(3H,m,H-2’+H-4’+Trp4-α-CH),3.90(1H,m,DABA1-β-CH),3.71- 3.50(4H,m,Gly2-α-CH2+Trp4-β-CH2),3.24–2.99(4H,m,Trp4-β-CH2+2x H-5’),2.73(3H,s, NCH3),2.06–1.98(1H,m,H-3’),1.76–1.65(1H,m,H-3’),1.13(3H,m,DABA1-γ-CH3).1H NMR (400MHz,DMSO-d6, the ratio 1.3:1 of rotational isomer, rotational isomer 2): δ 2.17-2.08 (1H, m, H-3 ')13C NMR(101MHz,DMSO-d6；Rotamer ratio 1.3:1, rotational isomer 1): δ 175.1 (C=O), 174.8 (C=O), 173.0 (C=O), 169.9 (C=O), 168.7 (C=O), 167.9 (C=O), 163.6,150.8 (C=O), 141.5, 136.2,128.5,128.0,127.7,123.8,121.0,118.9,118.5,111.5,110.6,101.9,92.4,78.7, 74.4,57.0,55.4,54.6,53.3,50.8,44.0,40.5,36.5,29.1,28.3,27.3,14.9.13C NMR (101MHz,DMSO-d6, the ratio 1.3:1 of rotational isomer, rotational isomer 2): δ 35.7.LRMS [M+H+]762.4.HRMS (ESI m/z)[M+H+]C36H44N9O10Calculated value 762.3206, measured value 762.3200.
(2S, 6S, 9S, 10S) -2- ((1H- indol-3-yl) methyl) -13- amino -9- ((((2R, 4R, 5R) -5- (2,4- - 1 (2H)-yl of dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) carbamoyl) -10,11- diformazan Four azepine tridecane -1- of base -4,7,12- trioxy- -6- phenethyl -3,5,8,11- is sour (35)
According to general step 8, at HOAt (23.9mg, 175.5 μm of ol), EDC.HCl (8.7mg, 45.6 μm of ol) and NMM In the presence of (5.0 μ L, 45.6 μm of ol), make depsipeptide S50 (29.3mg, 35.1 μm of ol) and amine 18 (19.9mg, 87.7 μm of ol) In CH2Cl2: reaction 4 hours in DMF (1:1v/v, 360 μ L) obtain shielded analog completely.Then with according to general side Method 8, the compound is with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.% iPr3SiH, 1.2mL) 16 hours, fluffy white solid 35 is obtained after reversed-phase HPLC purifies (0 to 50%MeCN, 40 minutes) (formates) (7.0mg, two steps, 24%).
IR(ATR):3377,2960,2929,1680,1435cm-1.1H NMR(500MHz,DMSO-d6, rotational isomer Ratio 1.5:1, main rotational isomer): δ 7.53 (2H, m, H-6+Ar-H), 7.30 (7H, m, 7x Ar-H), 7.01 (2H,m,2x Ar-H),5.64(1H,m,H-1’),5.55(1H,m,H-5),4.56(1H,m,DABA1-α-CH),4.28(3H, m,Trp4-α-CH+H-2’+H-4’),4.10–4.04(2H,m,HPhe3-α-CH+DABA1-β-CH),3.69(2H,m,Gly2- α-CH2),3.26-3.14(4H,m,Trp4-β-CH2+2x H-5’),2.73(3H,s,NCH3),2.50(2H,obscure, HPhe3-γ-CH2),2.09(1H,m,H-3’),1.83-1.66(3H,m,H-3’+HPhe3-β-CH2),1.12(3H,m, DABA1-γ-CH3).13C NMR(126MHz,DMSO-d6, the ratio 1.5:1 of rotational isomer, main rotational isomer): δ 177.7 (C=O), 172.7 (C=O), 171.4 (C=O), 169.9 (C=O), 169.0 (C=O), 163.7 (C=O), 150.8 (C=O), 141.4,136.3,128.8,128.7,126.2,126.0,125.1,124.1,121.0,11 8.9,111.4, 110.3,101.6,92.5,79.0,78.9,74.2,55.5,53.4,52.9,50.8,43.8,40.9,40.3,35.7,31.4, 29.2,27.5,15.1.LRMS[M+H+]790.5.HRMS(ESI m/z)[M+H+] calculated value C38H48N9O10790.3518, it surveys Magnitude 790.3509.
(((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- base) ammonia Base) -3- (naphthalene -2- base) -1- oxopropan -2- base) carbamoyl)-L-Trp (36)
According to the condition B of general step 8, at room temperature, HOAt (17mg, 123 μm of ol) and EDC.HCl (6.1mg, 32 μ Mol) and in the presence of NMM (3.5 μ L, 32 μm of ol), depsipeptide S51 (21mg, 25 μm of ol) and amine 18 (14mg, 63 μm of ol) is made to exist CH2Cl2: reaction 4 hours in DMF (1:1v μ/v, 250 μ L) obtain shielded analog completely.According to universal method 8, the change It closes object and with TFA and contains iPr3The CH of SiH2Cl2Mixture handles (1:1v/v TFA:CH2Cl2, 2.5vol.%iPr3SiH, 2.5mL) 4 hours, reversed-phase HPLC (0 to 50%MeCN, 40 minutes, 10 minutes, 100%H2O, flow velocity 9mL/min) after purification, Obtain amorphous white solid 36 (formates) (6.0mg, 28%, 2 step).
IR(ATR):3390,2977,2936,1730,1644cm-1.1H NMR(500MHz,CD3OD, main rotation are different Structure body): δ 8.56 (1H, t, J=6.0Hz, 1H), 8.16 (1H, d, J=9.1Hz), 8.13 (0.4H, d, J=8.6Hz), 7.84 (1H, t, J=5.9Hz), 7.81-7.69 (2H, m, Ar-H), 7.65-7.60 (1H, m, Ar-H), 7.58-7.54 (2H, m, Ar-H +H-6),7.45-7.40(3H,m,Ar-H),7.36-7.29(2H,m,Ar-H),7.11-7.04(2H,m,Ar-H),7.03- 6.92(1H,m,Ar-H),5.72-5.66(2H,m,H-1’+H-5),4.64-4.53(1H,m,Trp-α-CH),4.53-4.48 (1H,m,DABA1-α-CH),4.48-4.34(2H,H-2’+Nal3-α-CH),4.25-4.16(1H,m,H-4’),4.07(1H D, J=15.9Hz, Gly2- α-CH2),3.88-3.81(2H,m,DABA1-β-CH+Gly2-α-CH2),3.30-3.23(1H,m, Trp4-β-CH2),3.22-3.04(4H,m,H-5’+Trp4-β-CH2+Nal3-β-CH2),3.13-3.03(1H,m,H-5’), 2.91 (1H, ddd, J=13.6,8.6,4.8Hz, H-5 '), 2.83 (3H, s, NCH3), 2.14 (1H, ddd, J=13.6,7.5, 6.0Hz, H-3 '), and 1.66-1.59 (1H, m, H-3 '), 1.17 (3H, d, J=6.6Hz, DABA1- γ-CH3).13C NMR (125MHz,CD3OD, main rotational isomer, two13C signal is fuzzy): δ 176.0 (C=O), 174.4 (C=O), 171.0 (C=O), 167.6 (C=O), 166.3 (C=O), 159.5 (C=O), 152.4 (C=O), 142.6,142.4,137.6, 135.8,134.7,134.0,128.8(×2),128.6,128.2,126.6,124.4,122.0,119.6,119.2,112.0, 110.6,102.3,94.8,81.1,76.0,56.6(×2),55.0,54.8,45.0,40.9,39.2,35.9,29.1,27.7, 14.5.LRMS[M+H+]826.4.HRMS(ESI m/z)[M+H+]C41H48N9O10Calculated value 826.3518, measured value are 826.3515。
(((S) -1- (((2S, 3S) -3- ((S) -2- amino -3- (3- hydroxy phenyl)-N- methyl propanamide) -1- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) Amino) -1- oxo-butanes -2- base) amino) -3- cyclohexyl -1- oxopropan -2- base) carbamoyl)-L-Trp (37)
According to the condition B of general step 8, at room temperature, HOAt (11mg, 150 μm of ol) and EDC.HCl (4.4mg, 22 μ Mol) and in the presence of NMM (2.6 μ L, 22 μm of ol), depside ester S52 (18mg, 17 μm of ol) and amine 18 (12mg, 52 μm of ol) is made to exist CH2Cl2: sustained response 2.5 hours in DMF (1:1v/v, 180 μ L), obtain the analog protected completely.With TFA and contain iPr3The H of SiH2O (9:1v/v TFA:CH2Cl2, 2vol.%iPr3SiH, 1.7mL) in mixture handle the compound, 0 Keep 45 minutes at DEG C, 2 hours at room temperature, reversed-phase HPLC (0 to 50%MeCN, 40 minutes；10 minutes, 100%H2O) pure It is (8.0mg, 2 steps, yield 53%) that amorphous white solid 37 (tfa salt) is obtained after change.
IR(ATR):3388,2926,1647cm-1.1H NMR(500MHz,CD3OD, the ratio 2.6:1 of rotational isomer, Main rotational isomer): δ 7.62 (1H, d, J=8.1Hz, H-6), 7.54 (1H, app.t, J=7.8Hz, Ar-H), 7.32 (1H, app.d, J=8.2Hz, Ar-H), 7.18 (1H, app.t, J=7.9Hz, Ar-H), 7.12-7.04 (2H, m, Ar-H), 7.02-6.96 (1H, m, Ar-H), 6.78-6.71 (2H, m, Ar-H), 6.69-6.66 (1H, m, Ar-H), 5.74 (1H, d, J= 2.5Hz, H-1 '), 5.71 (1H, d, J=8.1Hz, H-5), 4.91-4.75 (1H, m, m-Tyr2- α-CH), 4.59-4.49 (2H, m,DABA1-α-CH+Trp4-α-CH),4.48-4.35(2H,H-2’+H-4’),4.23-4.07(2H,m,DABA1-β-CH+ Cha3- α-CH), 3.52 (1H, dd, J=13.9,3.9Hz, H-5 '), 3.29-3.23 (2H, m, H-5 '+Trp4- β-CH2), 3.23-3.11(1H,m,Trp4-β-CH2),3.02-2.97(2H,m,m-Tyr2-β-CH2),2.78(3H,s,NCH3),2.23 (1H, app.dt, J=13.7,7.0Hz, H-3 '), ('+2 5H, m, H-3 × CH 1.82-1.582),1.48-1.40(2H,m, Cha3-β-CH2),1.38-1.11(6H,m,Cha3-γ-CH+2.5×CH2),0.98-0.85(1H,m,0.5×CH2),0.82 (3H, d, J=6.5Hz, DABA1- γ-CH3).13C NMR(125MHz,CD3OD, the ratio 2.6:1 of rotational isomer, it is main Rotational isomer): δ 175.8 (C=O), 175.4 (C=O), 171.3 (C=O), 170.2 (C=O), 165.8 (C=O), 159.8 (C=O), 159.0,151.6 (C=O), 142.1,137.6,136.1,131.2,128.6,124.4,122.2, 121.7,119.7,119.3,116.9,115.7,112.1,110.5,102.3,94.9,80.9,76.2,56.9,54.6, 54.5,52.9,52.8,44.8,40.4,38.3,35.7,35.0,33.8,28.8,28.3,27.1,14.2.LRMS[M+H+] 888.3.HRMS(ESI m/z)[M+H+] calculated value C44H58N9O11888.4250 measured value 888.4244.
Tert-butyl (S) -3- (2- ((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino) -3- (allyloxy) -3- oxo Propyl) -1H- indoles -1- carboxylate (38)
At 0 DEG C, it is added dropwise into the solution of the DMF (2.8mL) of Fmoc-Trp (Boc)-OH (1.0g, 1.9mmol, 1eq.) iPr2Then allyl bromide, bromoallylene (330 μ L, 3.8mmol, 2eq.) is added dropwise in EtN (660 μ L, 3.8mmol, 2eq.).It is heated to reaction Room temperature, and stir 16 hours, then diluted with EtOAc (80mL).Then organic layer H2O (6 × 40mL), salt water (40mL) are washed It washs, dry (MgSO4) and be concentrated in vacuo, obtain white foam 38 (1.1g, 2.0mmol, quantitative).
IR(ATR):2923,2853,1731,1512cm-1； 1H NMR (500MHz,CDCl3)δ8.13-8.12(m,1H,Ar-H),7.76-7.75(m,2H,Ar-H),7.57-7.51(m,3H,Ar- H),7.41-7.37(m,3H,Ar-H),7.33-7.27(m,3H,Ar-H),7.24-7.21(m,1H,Ar-H),5.89-5.81 (m,1H,OCH2CH=CH2) 5.41 (d, J=8.4Hz, 1H, NH), 5.31-5.23 (m, 2H, OCH2CH=CH2),4.80-4.77 (m, 1H, α-CH), 4.59 (d, J=5.5Hz, 2H, OCH2CH=CH2),4.42-4.34(m,2H,Fmoc-CH2),4.21(t,J =7.1Hz, 1H, Fmoc-CH), 3.52 (d, J=4.6Hz, 2H, β-CH2),1.66(s,9H,CO2t-Bu)；13C NMR (125MHz,CDCl3)δ171.5,155.9,149.7,144.0,143.9,141.4,138.2,135.5,131.5,130.7, 128.9,127.9,127.2,127.2,125.3,124.8,124.4,122.8,122.8,121.2,120.1,120.0, 119.4,119.0,115.5,114.9,107.9,83.9,67.4,66.4,54.4,47.3,28.4,28.1；LRMS[M+Na]+ 589.4.
Tert-butyl (S) -3- (3- (allyloxy) -2- amino -3- oxopropyl) -1H- indoles -1- carboxylate (39)
Allyl ester 38 (1.1g, 2.0mmol, 1eq.) is handled with MeCN (9.5mL) solution of 20%v/v piperidines, and Stirring 30 minutes.Be removed in vacuum solvent, gained residue purified by column chromatography (eluent: 1:1v/v hexane: EtOAc, 0.1%v/v diethylamine is to pure EtOAc, 0.1%v/v diethylamine), obtain yellow oily 39 (0.56g, 1.6mmol, 85%).
IR(ATR):2924,2854,1732,1454cm-1； 1H NMR (400MHz,CDCl3)δ8.13-8.11(m,1H,Ar-H),7.57-7.55(m,1H,Ar-H),7.51(m,1H,Ar-H), 7.33-7.29(m,1H,Ar-H),7.25-7.22(m,1H,Ar-H),5.91-5.83(dddd,1H,OCH2CH=CH2),5.29 (dq, J=17.2,1.5Hz, 1H, OCH2CH=CH2), 5.23 (dq, J=10.4,1.2Hz, 1H, OCH2CH=CH2),4.59 (ddd, J=6.0,5.9,1.3Hz, 2H, OCH2CH=CH2), 3.94 (dd, J=7.5,5.3Hz, 1H, α-CH), 3.26 (dd, J =14.3,5.1Hz, 1H, β-CH2) 3.08 (dd, J=14.4,7.4Hz, 1H, β-CH2),1.66(s,9H,CO2t-Bu)；13C NMR(100MHz,CDCl3)δ174.7,149.6,135.5,131.8,130.4,124.5,124.2,122.5,118.9, 118.8,116.0,115.3,83.6,65.7,54.5,30.5,28.2；LRMS[M+H]+345.1.
Tert-butyl (S) -3- (3- (allyloxy) -2- (((4-nitrophenoxy) carbonyl) amino) -3- oxo-propyll) - 1H- indoles -1- carboxylate (40)
To 39 (0.56g, 1.6mmol, 1eq.) and iPr2The CH of EtN (2804-, 1.6mmol, 1eq.)2Cl2(8mL) solution The CH of middle dropwise addition p-nitrophenyl chloro-carbonic acid (0.33g, 1.6mmol, 1eq.)2Cl2(4mL) solution) and stir 1 hour.It is removed in vacuum Solvent obtains thick yellow oily 40 (1.3g), is directly used in subsequent step.
Tert-butyl (S) -3- (2- ((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino) -3- methoxyl group -3- oxo third Base) -1H-indoles -1- carboxylate (41)
At 0 DEG C, it is added dropwise into MeOH (3mL) solution of FmocTrp (Boc)-OH (0.20mg, 0.38mmol, 1eq.) SOCl2(40 μ L, 0.53mmol, 1.4eq.).Then heating is reacted to room temperature, and is stirred 30 minutes, then small in 40 DEG C of heating 4 When.Then it is cooled to room temperature reaction, is concentrated in vacuo.Thick residue is re-dissolved in MeOH (1.6mL), and vacuum again Concentration.Flask is cooled to 0 DEG C, residue is then dissolved in Na2CO3In the saturated aqueous solution of (5mL).By reactant CHCl3 (10mL) and H2O (5mL) extraction.By water phase CHCl3(10mL) extraction is primary.Combined organic extract liquid is dry (MgSO4), and be concentrated in vacuo.Gained foam is re-dissolved in CH2Cl2In, and be concentrated in vacuo again, canescence foam is obtained, with Afterwards by column chromatography eluting (eluent: 4:1v/v hexane: EtOAc), obtain white foam 41 (0.11mg, 0.20mmol, 52%).
IR(ATR):3333,2924,1730cm-1； 1H NMR(300MHz, CDCl3)δ8.12(m,1H,Ar-H),7.78-7.74(m,2H,Ar-H),7.58-7.49(m,3H,Ar-H),7.43-7.32(m, 7H,Ar-H),5.38(m,1H,NH),4.83-4.71(m,1H,α-CH),4.40-4.36(m,2H,Fmoc-CH2),4.28- 4.21(m,1H,Fmoc-CH),3.71(s,3H,CH3),3.28-3.26(m,2H,β-CH2),1.65(s,4.5H,CO2t-Bu), 1.57(s,4.5H,CO2t-Bu)；13C NMR(75.5MHz,CDCl3)δ172.1,155.7,149.5,143.7,141.2, 135.3,130.4,127.6,127.0,125.1,124.5,124.1,122.6,119.9,118.7,115.3,114.9,83.6, 67.1,52.3,47.0,28.1,27.7；LRMS[M+Na]+563.5；HRMS calculated value C32H32N2O6: MNa+, 563.21581.It surveys Magnitude: MNa+,563.21531。
Tert-butyl (S) -3- (3- methoxyl group -2- (((4-nitrophenoxy) carbonyl) amino) -3- oxopropyl) -1H- Yin Diindyl -1- carboxylate (42)
Compound 41 (0.69g, 1.3mmol, 1eq.) is small with MeCN (6.5mL) the solution processing 1 of 20%v/v piperidines When.By column chromatography eluting crude mixture (eluent: 2:1v/v hexane: EtOAc, 0.1%v/v diethylamine to pure EtOAc, 0.1%v/v diethylamine), obtain yellow oil (0.27g).It is re-dissolved in CH2Cl2In (6.6mL), in iPr2EtN In the presence of (0.15mL, 0.86mmol, 1eq.), handled with p-nitrophenyl chloro-formate (0.21g, 1mmol, 1.2eq.), and Stirring 20 hours, obtains carbamate.By column chromatography (eluent: 3:1v/v hexane: EtOAc), white foam is obtained (0.37g, 0.77mmol, 2 step, 60%).
IR(ATR):3346,2923,1733,1595,1371cm-1； 1H NMR(300MHz,CDCl3)δ8.23-8.20(m,2H,Ar-H),8.15-8.12(1H,Ar-H),7.55-7.49(m,2H,Ar- H),7.37-7.32(m,1H,Ar-H),7.24(m,3H,Ar-H),5.91(m,1H,NH),4.79-4.77(m,1H,α-CH), 3.78(s,3H,CH3),3.41-3.27(m,2H,β-CH2),1.69(s,9H,CO2t-Bu)；13C NMR(75.5MHz,CDCl3)δ 171.6,155.6,152.7,149.6,144.9,135.4,130.4,125.1,124.8,124.3,122.7,122.0, 118.7,115.5,114.5,84.0,54.4,52.8,28.2；LRMS[M+Na]+506.13；HRMS calculated value: MNa+, 506.15393.Measured value: MNa+,506.15349。
3- ethyoxyl -3- oxopropanoic acid (43)
At 0 DEG C, to the 1:10v/v THF:H of diethyl malonate (3.1mL, 20mmol, 1eq.)2O (400mL) solution Middle dropwise addition KOH(aq)(64mL, 0.25M, 16mmol, 0.8eq.).Reactant is stirred 1 hour, then at 0 DEG C, uses 1M HCl(aq)It is acidified to pH 2.Then by aqueous solution saturation NaCl aqueous solution (50mL) dilution, extracted with EtOAc (3 × 200mL) It takes, dry (MgSO4) and be concentrated in vacuo.Gained residue purifies (2:1v/v hexane: EtOAc) by column chromatography, obtains colourless Oily 43 (1.2g, 8.9mmol, 45%).
IR(ATR):3000,1714cm-1；1H NMR(300MHz,CDCl3) δ 4.24 (q, J=7.1Hz, 2H, OCH2), 3.43(s,2H,CH2), 1.30 (t, J=7.2Hz, 3H, CH3)；13C NMR(75.5MHz,CDCl3)δ170.6,167.2,62.0, 40.6,14.0；LRMS[M+H]+133.1.These data and Niwayama et al. in Tetrahedron Lett2008,49, 4434-4436. is reported consistent.
Monoethyl malonate magnesium (44)
Compound 43 is added in anhydrous THF (11mL) solution of magnesium ethylate (0.47g, 4.1mmol, 1eq.), and is stirred It mixes 2 hours.Obtained yellow solution is concentrated in vacuo, and is used for subsequent step, it is not necessary that the reorganization of the step is further purified certainly Kuhn, S. et al. are published in J.Am.Chem.Soc.2011,133,3708-3711. step 1- (tert-butyl) -6- ethyl (S) - 2- ((tertbutyloxycarbonyl) amino) -4- oxo adipate ester (45)
Carbonyl dimidazoles are added into anhydrous THF (30mL) solution of Boc-Asp-OtBu (2.2g, 7.5mmol, 1eq.) It (1.3g, 8.3mmol, 1.1eq.) and stirs 2 hours.Then the reaction is cooled to 0 DEG C, then by 44 anhydrous THF (5.2mL) Above-mentioned solution is added in solution.Then heating reaction to room temperature and is stirred 20 hours, then uses H2O (30mL) dilution, and use 1M HCl (aqueous solution) is acidified to pH 2.Then aqueous solution Et2O (3 × 30mL) extraction.Organic phase saturation NaHCO3Aqueous solution, NaHCO3(aqueous solution) (30mL) washing, dry (MgSO4) and be concentrated in vacuo.Gained residue purifies (2:1v/v by column chromatography Hexane: EtOAc), obtain yellow oily 45 (1.9g, 5.3mmol, 71%).
IR(ATR):2979,2923,2852,1718cm-1； 1H NMR (500MHz,CDCl3) δ 5.41 (d, J=8.1Hz, 1H, NH), 4.40-4.37 (m, 1H, α-CH), 4.19 (q, J=7.2Hz, 2H,OCH2),3.44(s,2H,CH2), 3.21 (dd, J=18.2,4.4Hz, 1H, β-CH2), 3.04 (dd, J=18.1,4.3Hz, 1H,β-CH2)；1.43(s,18H,9x CO2T-Bu+9x Ot-Bu), 1.27 (t, J=7.1Hz, 3H, CH3)；13C NMR (75.5MHz,CDCl3)δ200.7,169.9,166.5,155.5,82.2,79.8,61.4,50.0,49.2,45.0,28.3, 27.8,14.0；LRMS[M+Na]+382.2.These data and Brun, M. et al. are published in Angew.Chem.Int.Ed.Engl.2004,43,3432-3436. the data on are consistent.
(S) -1- carboxyl -2- (6,7- dimethoxy -2- oxo -2H- chromene -3- base) second -1- amino methyl sulfonic acid Salt (46)
At room temperature, with 1:1v/v TFA:CH2Cl2It is small that (14mL) handles (1.5g, 4.2mmol, the 1eq.) 1 of compound 45 When.Then be removed in vacuum solvent and with toluene (3 × 20mL) azeotropic, be then added 3,4- syringol (0.97g, 6.3mmol, 1.5eq.).Then it at 0 DEG C, is added methanesulfonic acid (6.8mL, 100mmol, 25eq.), is then heated to reaction Room temperature simultaneously stirs 2 hours.Product is precipitated out in ether, and with 3600g centrifugation 20 minutes.Sediment is re-dissolved in H2It in O and is lyophilized, obtains blue oily 46 (3.2g, 8.1mmol, quantitative).
IR(ATR):2954,2923,1707,1613cm-1； 1H NMR (300MHz,(CH3)2SO)δ8.42(s,3H,NH3),7.15-7.06(m,2H,Ar-H),6.28(s,1H,H-3),4.30-4.25 (m, 1H, α-CH), 3.84 (s, 3H, OMe), 3.83 (s, 3H, OMe), 3.36 (dd, J=14.7,6.4Hz, 1H, β-CH2), 3.26 (dd, J=14.7,8.0Hz, 1H, β-CH2)；13C NMRδ173.0,160.7,156.4,153.3,149.5,146.4, 112.5,111.5,106.2,100.9,56.6,56.5,53.4,47.0,33.3；LRMS[M+H]+294.0.These data with Brun, M. et al. are published in Angew.Chem.Int.Ed.Engl.2004, and the data on 43,3432-3436. are consistent.
(S) -2- ((((9H- fluorenes -9- base) methoxyl group) carbonyl) amino) -3- (6,7- dimethoxy -2- oxo -2H- benzo Pyrans -3- base) propionic acid (47)
To the 1.5:1v/v THF of 46 (3.0g, 7.7mmol, 1eq.): saturation NaHCO3(aq)It is added in (77mL) solution Fmoc-OSu (2.8mg, 8.4mmol, 1.1eq.) is simultaneously stirred 17 hours.Then by reaction H2O (80mL) dilution, and use ether (3 × 80mL) washing.Then water phase is acidified to pH 2 with 1M HCl (aqueous solution), is extracted with EtOAc (3 × 100mL), it is dry (MgSO4) and be concentrated in vacuo, obtain green powder 25 (2.0g, 3.8mmol, 50%).
IR(ATR):3368,1701,1614cm-1； 1H NMR(500MHz, (CH3)2SO)δ7.88-7.86(m,2H,Ar-H),7.63-7.60(m,2H,Ar-H),7.40-7.38(m,2H,Ar-H),7.29- 7.25(m,3H,Ar-H),7.07(m,1H,Ar-H),6.24(s,1H,H-3),4.37-4.32(m,1H,α-CH),4.24-4.22 (m,2H,Fmoc-CH2), 4.16 (m, 1H, Fmoc-CH), 3.85 (s, 3H, OMe), 3.84 (s, 3H, OMe), 3.32 (dd, J= 14.5,4.0Hz,1H,β-CH2) 3.10 (dd, J=14.4,9.9Hz, 1H, β-CH2)；13C NMRδ173.0,160.7,156.4, 153.3,149.5,146.4,144.2,144.1,144.1,144.1,128.1,127.5,125.6,125.6,120.6, 112.5,111.5,106.2,100.9,66.2,56.6,56.5,53.4,47.0,33.3,25.7；LRMS[M+H]+516.3.This A little data and Brun, M. et al. are published in Angew.Chem.Int.Ed.Engl.2004, the data one on 43,3432-3436. It causes.
General step 7.1
In CH2Cl2In (0.04M), by the false peptide of resin-bonded with tetrakis triphenylphosphine palladium (0) (0.028-0.052mmol, 0.20eq.) solution and phenyl silane (2.8-5.2mmol, 20eq.) processing, while shake 15 minutes, then with DMF (5 × 5mL), CH2Cl2(5 × 5mL) and DMF (5 × 5mL) wash resin.A small amount of cracked solution, which is detected, by LCMS confirms that Alloc- is protected Shield base completely removes.
General step 7.2
Condition A:
In anhydrous CH2Cl2In (0.5M), false peptide 51-54,58-62 N, N '-diisopropylcarbodiimide of resin-bonded (DIC) (0.62-1.3mmol, 5eq.) is vibrated 5 minutes and is activated.Then alcohol (0.62-1.3mmol, 5eq.) and 4- bis- is wherein added The anhydrous CH of dimethylaminopyridine (DMAP) (12-26 μm of ol, 0.1eq.)2Cl2(0.5M) solution.Resin oscillation 16-20 is small When, DMF (5 × 5mL) then is used, CH2Cl2(5 × 5mL) and DMF (5 × 5mL) washing.Pass through the LCMS to a small amount of cracked solution Detection is coupled successfully.
Condition B:
In anhydrous CH2Cl2In (0.5M), false peptide 55-57,63-65 N, the N- diisopropylcarbodiimide of resin-bonded (DIC) (0.54-0.98mmol, 5eq.) is vibrated 5 minutes and is activated.The anhydrous of amine (0.21-0.39mmol, 2eq.) is added thereto CH2Cl2(0.5M) solution.Resin is shaken 2 hours, then uses DMF (5 × 5mL), CH2Cl2(5 × 5mL) and DMF (5 × 5mL) Washing.It is coupled successfully by carrying out LCMS detection confirmation to a small amount of cracked solution.
General step 8
Condition C:
Pass through the CH with 30%v/v HFIP2Cl2(5mL) solution is handled 40 minutes, is vibrated at room temperature, is supported from solid False peptide 51-65 is cracked on object.Then CH is used2Cl2(6 × 10mL) washs resin, merges with cracked solution and is concentrated in vacuo.Then Thick residue is directly used in molten liquid phase coupling without being further purified.
In 2:1v/v CH2Cl2: in DMF (190-260 μ L), with NMM (46-65Mmol, 1.3eq.) to EDCHCl (46- 65 μm of ol, 1.3eq.) it vibrates 15 minutes at room temperature, carry out pre-activate.Then at 0 DEG C, which is added drop-wise to thick residue 51- 65 (36-50 μm of ol, 1eq.), uracil amine 18 (0.11-0.15mmol, 3eq.) and HOAt's (0.18-0.25mmol, 5eq.) 1:2v/v CH2Cl2: in DMF (190-260 μ L) solution.So that reaction is heated to room temperature and is stirred 3-4 hours, then uses EtOAc (44-60mL) dilution is saturated NaHCO with 0.5M HCL aqueous solution (11-16mL)3Aqueous solution (11-16mL), H2O(11- 16mL), salt water (11-16mL) washs, and dry (MgSO4).Organic phase is concentrated in vacuo, then again with 1:1v/v CH2Cl2: TFA (5.2-6.2mL) and 2.5vol.%iPr3SiH (0.11-0.16mL) is redissolved.By reaction stirring 14-17 hours, then Vacuum concentration.Obtained residue is re-dissolved in the H of 1%TFA2In O (25mL), and in N2(g)Flow down blowout.Then gained is residual Excess is redissolved in the H containing 20-30%v/v MeCN2In O, and purified by reversed-phase HPLC.
Resin-bonded false peptide (48)
As described in general step 2, iPr will be contained2The CH of EtN (350 μ L, 2mmol, 4eq.)2Cl2In (0.1M, 5.5mL) Amino acid 20 (240mg, 0.55mmol, 1.1eq.) be loaded to 2- chlorine trityl (2-CTC) resin (420-560mg, 0.50mmol, 1eq.) on, Alloc deprotection in DMF (0.1M, 11mL), is used then as described in general step 5 and 6 HATU (420mg, 1.1mmol, 2eq.) and iPr2EtN (290 μ L, 1.7mmol, 3eq.) and Boc-Gly-OH (190mg, 1.1mmol, 2eq.) coupling.Subsequent Fmoc deprotection in DMF (0.1M, 12mL), is used according to 3 condition A of general step PyBOP (0.62g, 1.2mmol, 4eq.) and NMM (0.27mL, 2.4mmol, 8eq.) and FmocCha-OH (0.47g, 1.2mmol, 4eq.) coupling.IPr is used in anhydrous DMF (0.1M, 3.5mL) according to general step 42EtN (0.12mL, 0.70mmol, 4eq.), thick allyl carbamate 40 (0.18g, 0.35mmol, 2eq.) and the de-protected resin knot of Fmoc The tripeptides of conjunction is coupled.According to general step 7.1, allyl deprotection protection obtains the false peptide 48 of resin-bonded.
Resin-bonded false peptide (49)
As described in general step 2, iPr will be contained2The CH of EtN (350 μ L, 2mmol, 4eq.)2Cl2In (0.1M, 5.5mL) Amino acid 20 (240mg, 0.55mmol, 1.1eq.) be loaded to 2- chlorine trityl (2-CTC) resin (420-560mg, 0.50mmol, 1eq.) on, Alloc deprotection in DMF (0.1M, 11mL), is used then as described in general step 5 and 6 HATU (420mg, 1.1mmol, 2eq.) and iPr2EtN (290 μ L, 1.7mmol, 3eq.) and Boc-Gly-OH (190mg, 1.1mmol, 2eq.) coupling.Subsequent Fmoc deprotection in DMF (0.1M, 12mL), is used according to 3 condition A of general step PyBOP (0.54g, 1.1mmol, 4eq.) and NMM (0.23mL, 2.1mmol, 8eq.) and Fmoc2-Nal-OH (0.46g, 1.1mmol, 4eq.) coupling.IPr is used in anhydrous DMF (0.1M, 3.5mL) according to general step 42EtN (0.12mL, 0.70mmol, 4eq.), thick allyl carbamate 40 (0.18g, 0.35mmol, 2eq.) and the de-protected resin knot of Fmoc The tripeptides of conjunction is coupled.According to general step 7.1, allyl deprotection protection obtains the false peptide 49 of resin-bonded.
The false peptide (50) of resin-bonded
As described in general step 2, iPr will be contained2The CH of EtN (350 μ L, 2mmol, 4eq.)2Cl2In (0.1M, 5.5mL) Amino acid 20 (240mg, 0.55mmol, 1.1eq.) be loaded to 2- chlorine trityl (2-CTC) resin (420-560mg, 0.50mmol, 1eq.) on, Alloc deprotection in DMF (0.1M, 11mL), is used then as described in general step 5 and 6 HATU (420mg, 1.1mmol, 2eq.) and iPr2EtN (290 μ L, 1.7mmol, 3eq.) and Boc-Gly-OH (190mg, 1.1mmol, 2eq.) coupling.Compound 47 (0.13g, 0.25mmol, 1.5eq.), HATU (95mg, 0.25mmol, 1.5eq.), HOAt (34mg, 0.25mmol, 1.5eq.) and iPr2The anhydrous DMF (0.1M, 1.7mL) of EtN (89 μ L, 0.51mmol, 3eq.) Solution is added in the peptide of the same race of the resin-bonded of Fmoc deprotection, and vibrates 2 hours.With DMF (5 × 5mL), CH2Cl2(5× 5mL) and DMF (5 × 5mL) washs resin, is then blocked, is vibrated 3 minutes with the pyridine (5mL) of 10%v/v acetic anhydride.Use DMF (5 × 5mL), CH2Cl2(5 × 5mL) and DMF (5 × 5mL) washing.According to general step 4, at anhydrous DMF (0.1M, 3.5mL) In, use iPr2EtN (0.12mL, 0.70mmol, 4eq.), thick allyl carbamate 40 (0.18g, 0.35mmol, 2eq.) It is coupled with the tripeptides of the de-protected resin-bonded of Fmoc.Then according to 7.2 condition A of general step, the false peptide of resin-bonded is used DIC (150 μ L, 0.96mmol, 5eq.) is in CH2Cl2Pre-activate in (1.9mL), then be added benzylalcohol (99 μ L, 0.96mmol, 5eq.) and the CH of DMAP (2.3mg, 19 μm of ol, 0.1eq.)2Cl2In (1.9mL) solution, the false peptide 50 of resin-bonded is obtained.
Resin-bonded false peptide (51)
According to 7.2 condition A of general step, the false peptide 48 of resin-bonded is existed with DIC (150 μ L, 0.97mmol, 5eq.) CH2Cl2Pre-activate in (1.9mL), be then added methanol (39 μ L, 0.97mmol, 5eq.) and DMAP (2.4mg, 19mol, CH 0.1eq.)2Cl2(1.9mL) solution obtains the false peptide 51 of resin-bonded.
Resin-bonded false peptide (52)
According to 7.2 condition A of general step, the false peptide 48 of resin-bonded is existed with DIC (140 μ L, 0.88mmol, 5eq.) CH2Cl2Pre-activate in (1.8mL), be then added hexanol (110 μ L, 0.88mmol, 5 μ eq.) and DMAP (2.14mg, 17.5mol, CH 0.1eq.)2Cl2(1.74mL) solution obtains the false peptide 52 of resin-bonded.
Resin-bonded false peptide (53)
According to 7.2 condition A of general step, the false peptide 48 of resin-bonded is existed with DIC (110 μ L, 0.69mmol, 5eq.) CH2Cl2Dodecanol (150, so, 0.69mmol, 5eq.) and DMAP (1.7mg, 14 μ is then added in pre-activate in (1.4mL) Mol, 0.1eq.) CH2Cl2(1.4mL) solution obtains the false peptide 53 of resin-bonded.
Resin-bonded false peptide (54)
According to 7.2 condition A of general step, the false peptide 48 of resin-bonded is existed with DIC (96 μ L, 0.62mmol, 5eq.) CH2Cl2Pre-activate in (1.2mL), be then added benzyl alcohol (64 μ L, 0.62mmol, 5eq.) and DMAP (1.5mg, 12 μm of ol, CH 0.1eq.)2Cl2(1.2mL) solution obtains the false peptide 54 of resin-bonded.
Resin-bonded false peptide (55)
According to 7.2 condition B of general step, the false peptide 48 of resin-bonded is existed with DIC (150 μ L, 0.98mmol, 5eq.) CH2Cl2The CH of hexylamine (52 μ L, 0.39mmol, 2eq.) is then added in pre-activate in (2mL)2Cl2(2mL) solution, obtains resin In conjunction with false peptide 55.
Resin-bonded false peptide (56)
According to 7.2 condition B of general step, the false peptide 48 of resin-bonded is existed with DIC (83 μ L, 0.54mmol, 5eq.) CH2Cl2The CH of neopentyl amine (25 μ L, 0.21mmol, 2eq.) is then added in pre-activate in (1.1mL)2Cl2(1.1mL) solution, obtains To the false peptide 56 of resin-bonded.
Resin-bonded false peptide (57)
According to 7.2 condition B of general step, the false peptide 48 of resin-bonded is existed with DIC (90 μ L, 0.58mmol, 5eq.) CH2Cl2The CH of benzylamine (25 μ L, 0.23mmol, 2eq.) is then added in pre-activate in (1.2mL)2Cl2(1.2mL) solution, obtains The false peptide 57 of resin-bonded.
Resin-bonded false peptide (58)
As described in general step 2, iPr will be contained2The CH of EtN (350 μ L, 2mmol, 4eq.)2Cl2In (0.1M, 5.5mL) Amino acid 20 (240mg, 0.55mmol, 1.1eq.) be loaded to 2- chlorine trityl (2-CTC) resin (420-560mg, 0.50mmol, 1eq.) on, Alloc deprotection in DMF (0.1M, 11mL), is used then as described in general step 5 and 6 HATU (420mg, 1.1mmol, 2eq.) and iPr2EtN (290 μ L, 1.7mmol, 3eq.) and BocGly-OH (190mg, 1.1mmol, 2eq.) coupling.Then Fmoc deprotection in DMF (0.1M, 11mL), is used according to 3 condition A of general step PyBOP (0.54g, 1.1mmol, 4eq.) and NMM (0.23mL, 2.1mmol, 8eq.) and Fmoc2-Nal-OH (0.46g, 1.1mmol, 4eq.) coupling.IPr is used in anhydrous DMF (0.1M, 1.6mL) according to general step 42EtN (60 μ L, 0.35mmol, 4eq.), allyl carbamate 42 (78mg, 0.16mmol, 1.9eq.) and the de-protected resin-bonded of Fmoc Tripeptides coupling, obtain the false peptide 58 of resin-bonded.
Resin-bonded false peptide (59)
According to 7.2 condition A of general step, the false peptide 49 of resin-bonded is existed with DIC (190 μ L, 1.3mmol, 5eq.) CH2Cl2Pre-activate in (2.5mL), be then added hexanol (160 μ L, 1.3mmol, 5eq.) and DMAP (3.1mg, 25 μm of ol, CH 0.1eq.)2Cl2In (2.5mL), the false peptide 59 of resin-bonded is obtained.
Resin-bonded false peptide (60)
According to 7.2 condition A of general step, the false peptide 49 of resin-bonded is existed with DIC (200 μ L, 1.3mmol, 5eq.) CH2Cl2Pre-activate in (2.6mL), be then added dodecanol (290 μ L, 1.3mmol, 5eq.) and DMAP (3.2mg, 26 μm of ol, CH 0.1eq.)2Cl2In (2.6mL), the false peptide 60. of resin-bonded is obtained
Resin-bonded false peptide (61)
According to 7.2 condition A of general step, the false peptide 49 of resin-bonded is existed with DIC (130 μ L, 0.81mmol, 5eq.) CH2Cl2Pre-activate in (1.6mL), be then added neopentyl alcohol (88 μ L, 0.81mmol, 5eq.) and DMAP (2.0mg, 16 μm of ol, CH 0.1eq.)2Cl2In (1.6mL), the false peptide 61 of resin-bonded is obtained.
Resin-bonded false peptide (62)
According to 7.2 condition A of general step, the false peptide 49 of resin-bonded is existed with DIC (160 μ L, 1.1mmol, 5eq.) CH2Cl2Pre-activate in (2.1mL), be then added benzyl alcohol (110 μ L, 1.1mmol, 5eq.) and DMAP (2.6mg, 21 μm of ol, 0.1eq.).According to 7.2 condition A of general step, in CH2Cl2In (2.1mL), the false peptide 62 of resin-bonded is obtained.
Resin-bonded false peptide (63)
According to 7.2 condition B of general step, the false peptide 49 of resin-bonded is existed with DIC (150 μ L, 0.98mmol, 5eq.) CH2Cl2The CH of hexylamine (52 μ L, 0.39mmol, 2eq.) is then added in pre-activate in (2mL)2Cl2(2mL) solution, obtains resin In conjunction with false peptide 63.
Resin-bonded false peptide (64)
According to 7.2 condition B of general step, the false peptide 49 of resin-bonded is existed with DIC (85 μ L, 0.55mmol, 5eq.) CH2Cl2The CH of neopentyl amine (26 μ L, 0.22mmol, 2eq.) is then added in pre-activate in (1.1mL)2Cl2(1.1mL) solution, obtains To the false peptide 64 of resin-bonded.
Resin-bonded false peptide (65)
According to 7.2 condition B of general step, the false peptide 49 of resin-bonded is existed with DIC (130 μ L, 0.8mmol, 5eq.) CH2Cl2The CH of benzylamine (40 μ L, 0.32mmol, 2eq.) is then added in pre-activate in (1.6mL)2Cl2(1.6mL) solution, obtains The false peptide 65 of resin-bonded.
Methyl ((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- Base) amino) -3- cyclohexyl -1- oxopropan -2- base) carbamoyl)-L-Trp salt (66)
According to 8 condition C of general step, false peptide 51 is cracked from resin.By preactivated EDCHCl (10mg, 52 μ Mol, 1.3eq.) and NMM (5.7 μ L, 52 μm of ol, 1.3eq.) in 2:1v/v CH2Cl2: the solution of DMF (210 μ L) is added dropwise To thick residue 51 (32mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μ Mol, 5eq.) 1:2v/v CH2Cl2: in DMF (210 μ L) solution, the analog 66 protected.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer of 98:2 Mixture.It is purified by reversed-phase HPLC, obtaining white fluffy solid 66, (8.0mg, 8.8 μm ol, 22%, HPLC after purification with TFA The yield that salt calculates).
IR(ATR):3279,2926,1680cm-1；1H NMR(500MHz,(CD3)2The 1:1 of SO, ca. rotational isomer is mixed Close object) δ 11.29 (s, 1H, uracil-NH), 10.90 (s, 1H, Trp-NH), 8.56 (t, J=5.5Hz, 0.5H, CONHCH2), 8.23 (d, J=9.0Hz, 0.5H, CONH), 8.17 (d, J=9.1Hz, 0.5H, CONH), 8.08 (t, J=5.3Hz, 0.5H, CONHCH2),8.02(br s,3H,NH3), 7.53 (d, J=8.1Hz, 0.5H, H-6), 7.50 (d, J=8.1Hz, 0.5H, H- 6),7.45-7.43(m,1H,Ar-H),7.34-7.33(m,1H,Ar-H),7.12-7.11(m,1H,Ar-H),7.07-7.04 (m, 1H, Ar-H), 6.99-6.96 (m, 1H, Ar-H), 6.38 (d, J=8.0Hz, 1H, NHCONH), 6.34-6.31 (m, 1H, NHCONH), 5.71 (d, J=3.4Hz, 0.5H, H-1 '), 5.66 (d, J=3.4Hz, 0.5H, H-1 '), 5.61-5.58 (m, 1H, ), H-5 4.82-4.76 (m, 0.5H, DABA- β-CH), 4.58 (t, J=9.2Hz, 0.5H, DABA- α-CH), 4.49 (t, J= 9.2Hz,0.5H,DABA-α-CH),4.42-4.36(m,1.5H,1x Trp-α-CH+0.5x H-4’),4.35-4.30(m,1H, H-2’),4.28-4.25(m,0.5H,H-4’),4.16-4.12(m,1H,Cha-α-CH),3.94-3.77(m,2H,1.5x Gly-α-CH2+0.5x DABA-β-CH),3.59-3.57(m,0.5H,Gly-α-CH2),3.55(s,3H,OCH3),3.28- 3.22(m,1H,H-5’),3.18-3.13(m,1H,H-5’),3.06-3.05(m,2H,Trp-β-CH2),2.77(s,1.5H, NCH3),2.75(s,1.5H,NCH3),2.21-2.10(m,1H,H-3’),1.69-1.60(m,6H,1x H-3’+5x Cy), 1.41-1.28(m,2H,Cha-β-CH2),1.23(m,1H,Cy),1.15-1.08(m,3.5H,1.5x CH3+2x Cy),1.02 (d, J=7.1Hz, 1.5H, CH3),0.87-0.76(m,3H,Cy)；13C NMR(125MHz,(CD3)2SO, ca. rotational isomer 1:1 mixture) δ 173.8,173.5,170.4,170.0,166.7,166.3,163.9,163.8,158.6,15 8.4, 158.1,157.7,157.6,151.2,141.7,141.5,136.7,127.8,124.5,121.6,119.0,118.7, 112.0,109.6,102.3,102.0,92.6,92.5,79.2,74.6,74.5,55.4,55.0,54.2,53.4,52.2, 51.7,51.0,44.4,44.3,36.2,36.0,34.1,33.8,32.8,32.7,29.4,28.3,27.8,26.6,26.5, 26.3,15.2,14.4；LRMS[M+H]+796.7；HRMS calculated value C38H53N9O10: MH+,796.39882.Measured value: MH+, 796.39969。
Hexyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- Base) amino) -3- cyclohexyl -1- oxopropan -2- base) carbamoyl)-L-Trp salt (67)
According to 8 condition C of general step, false peptide 52 is cracked from resin.EDCHCl (8.8mg, 46 μm of ol, 1.3eq.) With the 2:1v/v CH of NMM (5.1 μ L, 46 μm of ol, 1.3eq.)2Cl2: DMF (180 μ L) solution pre-activate is added dropwise to thick residual Excess 52 (30mg, 35 μm of ol, 1eq.), uridine amine 18 (24mg, 110 μm of ol, 3eq.) and HOAt (24mg, 180 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (180 μ L) solution, obtain shielded analog 67.With 1:1v/v CH2Cl2: TFA (4.4mL) and 2.5vol.%iPr3SiH (0.11mL) processing deprotection obtains the separable diastereoisomer mixing of 98:2 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 67, (9.3mg, 9.5 μm ol, 27%, HPLC after purification in terms of tfa salt The yield of calculation).
IR(ATR):3306,2925,2854,1664,1555cm-1；(500MHz,(CD3)2SO, ca. rotational isomer 1:1 mixture) δ 11.29 (s, 1H, uracil-NH), 10.89 (s, 1H, Trp-NH), 8.56 (t, J=5.5Hz, 0.5H, CONHCH2), 8.23 (d, J=9.0Hz, 0.5H, CONH), 8.18 (d, J=9.1Hz, 0.5H, CONH), 8.08 (t, J= 5.3Hz,0.5H,CONHCH2),8.01(br s,3H,NH3), 7.53 (d, J=8.1Hz, 0.5H, H-6), 7.50 (d, J= 8.1Hz,0.5H,H-6),7.45-7.44(m,1H,Ar-H),7.34-7.32(m,1H,Ar-H),7.12-7.11(m,1H,Ar- ), H 7.07-7.04 (m, 1H, Ar-H), 6.98-6.95 (m, 1H, Ar-H), 6.37 (d, J=8.0Hz, 1H, NHCONH), 6.35- 6.32 (m, 1H, NHCONH), 5.70 (d, J=3.5Hz, 0.5H, H-1 '), 5.66 (d, J=3.5Hz, 0.5H, H-1 '), 5.61- 5.58 (m, 1H, H-5), 4.82-4.76 (m, 0.5H, DABA- β-CH), 4.59 (t, J=9.2Hz, 0.5H, DABA- α-CH), 4.49 (t, J=9.2Hz, 0.5H, DABA- α-CH), 4.41-4.36 (m, 1.5H, 1x Trp- α-CH+0.5x H-4 '), 4.35- 4.30(m,1H,H-2’),4.28-4.25(m,0.5H,H-4’),4.16-4.12(m,1H,Cha-α-CH),3.92-3.77(m, 4H,1.5x Gly-α-CH2+0.5x DABA-β-CH+2x OCH2(CH2)4CH3),3.59-3.54(m,0.5H,Gly-α-CH2), 3.28-3.21 (m, 1H, H-5 '), and 3.18-3.13 (m, 1H, H-5 '), 3.04 (d, J=6.4Hz, 2H, Trp- β-CH2),2.77 (s,1.5H,NCH3),2.75(s,1.5H,NCH3),2.21-2.10(m,1H,H-3’),1.69-1.60(m,6H,1x H-3’+5x Cy),1.44-1.38(m,2H,OCH2(CH2)4CH3),1.35-1.31(m,2H,Cha-β-CH2),1.24-1.08(m,10.5H, 3x Cy+1.5CH3,6x OCH2(CH2)4CH3), 1.02 (d, J=6.8Hz, 1.5H, CH3),0.87-0.77(m,6H,3x Cy+ 3x OCH2(CH2)4C H3)；13C NMR(125MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.5, 173.4,170.4,170.0,166.7,166.3,163.9,163.8,158.8,158.6,158.4,158.1,157.7, 157.6,151.2,141.7,141.5,136.7,127.8,124.3,121.5,119.0,118.7,116.6,112.0, 109.7,102.3,102.0,92.6,92.5,79.2,74.6,74.5,64.8,55.4,55.0,54.3,53.3,51.7, 44.4,44.3,36.2,36.0,34.0,33.9,33.8,32.8,32.7,31.4,29.4,28.5,28.4,27.8,26.6, 26.5,26.3,25.5,22.5,15.2,14.5,14.4；LRMS[M+H]+866.8；HRMS calculated value C43H63N9O10: MH+, 866.47707.Measured value: MH+,866.47737。
Dodecyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) - 5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo fourth Alkane -2- base) amino) -3- cyclohexyl -1- oxopropan -2- base) carbamoyl)-L-Trp salt (68)
According to 8 condition C of general step, false peptide 53 is cracked from resin.Preactivated EDCHCl (9.3mg, 48 μ L, 1.3eq.) and the 2:1v/v CH of NMM (5.3 μ L, 48 μm of ol, 1.3eq.)2Cl2: DMF (190 μ L) solution is added dropwise to thick residual Excess 53 (35mg, 37 μm of ol, 1eq.), uridine amine 18 (25mg, 110 μm of ol, 3eq.) and HOAt (25mg, 190 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (190 μ L) solution, obtain shielded analog 68.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 97:3 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 68, (12mg, 12 μm ol, 29%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3310,2924,2853,1671,1556cm-1；1H NMR(400MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 11.30 (s, 1H, uracil-NH), 10.89 (s, 1H, Trp-NH), 8.57 (t, J=5.5Hz, 0.5H, CONHCH2), 8.24 (d, J=9.1Hz, 0.5H, CONH), 8.19 (d, J=9.1Hz, 0.5H, CONH), 8.09 (t, J= 5.1Hz,0.5H,CONHCH2),8.01(br s,3H,NH3), 7.53 (d, J=8.1Hz, 0.5H, H-6), 7.50 (d, J= 8.1Hz,0.5H,H-6),7.45-7.43(m,1H,Ar-H),7.34-7.32(m,1H,Ar-H)7.11-7.10(m,1H,Ar- H),7.07-7.03(m,1H,Ar-H),6.98-6.95(m,1H,Ar-H),6.56,(s,1H,OH),6.34-6.31(m,2H, NHCONH), 5.71 (d, J=3.4Hz, 0.5H, H-1 '), 5.66 (d, J=3.3Hz, 0.5H, H-1 '), 5.61-5.58 (m, 1H, ), H-5 4.82-4.75 (m, 0.5H, DABA- β-CH), 4.57 (t, J=9.5Hz, 0.5H, DABA- α-CH), 4.49 (t, J= 9.1Hz,0.5H,DABA-α-CH)4.41-4.35(m,1.5H,1x Trp-α-CH+0.5x H-4’),4.32-4.30(m,1H, H-2’),4.28-4.24(m,0.5H,H-4’)4.15-4.12(m,1H,Cha-α-CH),3.92-3.77(m,4H,1.5x Gly- α-CH2+2x OCH2(CH2)10CH3+0.5x DABA-β-CH),3.58-3.53(m,0.5H,Gly-α-CH2),3.26-3.22 (m, 1H, H-5 '), and 3.18-3.12 (m, 1H, H-5 '), 3.04 (d, J=6.3Hz, 2H, Trp- β-CH2),2.77(s,1.5H, NCH3),2.74(s,1.5H,NCH3),2.21-2.09(m,1H,H-3’),1.68-1.59(m,6H,1x H-3’+5x Cy), 1.45-1.38(m,2H,OCH2(CH2)10CH3),1.36-1.28(m,2H,Cha-β-CH2),1.26-1.15(m,17H,1x Cy+ 16x OCH2(CH2)10CH3),1.12-1.10(m,5.5H,1.5x CH3+2x Cy+2x OCH2(CH2)10CH3), 1.01 (d, J= 6.8Hz,1.5H,CH3),0.86-0.78(m,6H,3x OCH2(CH2)10CH3+3x Cy)；13C NMR(125MHz,(CD3)2SO, Ca. the 1:1 mixture of rotational isomer) δ 173.5,173.4,170.4,170.0,166.7,166.3,163.9,163.8, 158.9,158.6,158.4,158.1,157.7,157.6,151.2,141.7,141.5,136.7,127.8,124.4, 121.5,118.9,118.7,116.6,112.0,109.7,102.3,102.0,92.6,92.5,79.2,74.6,74.5, 64.8,55.4,55.0,54.3,53.3,51.7,51.0,44.4,44.3,36.2,36.0,34.1,33.9,33.8,32.9, 32.8,31.9,29.6,29.6,29.6,29.5,29.3,29.2,28.6,28.5,26.6,26.5,26.3,25.8,22.7, 15.1,14.6；LRMS[M+H]+951.0；HRMS C49H75N9O10Calculated value: MH+,950.57097.Measured value: MH+, 950.57081。
Benzyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- Base) amino) -3- cyclohexyl -1- oxopropan -2- base) carbamoyl)-L-Trp salt (69)
According to 8 condition C of general step, false peptide 54 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 54 (35mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 69.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 97:3 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 69, (12mg, 12 μm ol, 29%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3316,2926,2851,1665,1552cm-1；1H NMR(400MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 11.29 (s, 1H, uracil-NH), 10.90 (s, 1H, Trp-NH), 8.55 (t, J=5.2Hz, 0.5H, CONHCH2), 8.24 (d, J=9.0Hz, 0.5H, CONH), 8.19 (d, J=8.9Hz, 0.5H, CONH), 8.07 (t, J= 5.4Hz,0.5H,CONHCH2),8.00(br s,3H,NH3), 7.53 (d, J=8.1Hz, 0.5H, H-6), 7.50 (d, J= 8.1Hz,0.5H,H-6),7.49-7.44(m,1H,Ar-H),7.36-7.34(m,1H,Ar-H),7.32-7.30(m,3H,Ar- H)7.19-7.17(m,2H,Ar-H),7.10-7.09(m,1H,Ar-H),7.07-7.05(m,1H,Ar-H),6.99-6.95(m, 1H, Ar-H), 6.40-6.36 (m, 2H, NHCONH), 5.71 (d, J=3.4Hz, 0.5H, H-1 '), 5.66 (d, J=3.6Hz, 0.5H, H-1 '), 5.61-5.57 (m, 1H, H-5), 5.00 (q, J=12.7Hz, 2H, OCH2(C6H5)),4.82-4.75(m, 0.5H, DABA- β-CH), 4.58 (t, J=9.6Hz, 0.5H, DABA- α-CH), 4.52-4.43 (m, 1.5H, 0.5x DABA- α- CH+1x Trp-α-CH)4.40-4.36(m,0.5H,H-4’),4.33-4.31(m,1H,H-2’),4.29-4.24(m,0.5H, H-4’)4.18-4.13(m,1H,Cha-α-CH),3.95-3.76(m,2H,1.5x Gly-α-CH2+0.5x DABA-β-CH), 3.59-3.53(m,0.5H,Gly-α-CH2),3.29-3.21(m,1H,H-5’),3.19-3.12(m,1H,H-5’),3.08(d, J=6.4Hz, 2H, Trp- β-CH2),2.77(s,1.5H,NCH3),2.74(s,1.5H,NCH3),2.21-2.09(m,1H,H- 3’),1.69-1.60(m,6H,1x H-3’+5x Cy),1.39-1.29(m,2H,Cha-β-CH2),1.28-1.22(m,1H, Cy),1.13-1.07(m,3.5H,2x Cy+1.5x CH3), 1.02 (d, J=6.9Hz, 1.5H, CH3),0.87-0.78(m,3H, Cy)；13C NMR(100MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.3,173.2,170.3,169.9, 166.5,166.2,163.7,163.7,158.5,158.1,157.6,157.5,151.0,141.5,141.3,136.6, 136.3,128.8,128.4,128.1,127.6,124.3,121.4,118.9,118.6,111.9,109.5,102.2, 101.9,92.5,92.3,79.0,74.5,74.4,66.2,55.2,54.9,54.3,53.2,51.6,44.2,44.2,36.0, 35.9,33.9,33.6,32.7,32.6,29.2,28.3,27.7,26.5,26.3,26.1,15.0,14.3；LRMS[M+H]+ 872.8；HRMS calculated value C44H57N9O10: MH+,872.43012.Measured value: MH+,872.43013。
(2S, 3S) -3- (2- Amino-N-methyl acetylamino) -2- ((S) -3- cyclohexyl -2- (3- ((S) -1- (hexyl ammonia Base) -3- (1H- indol-3-yl) -1- oxo propyl- 2- yl) urea groups) propionamido)-N- (((2R, 4R, 5R) -5- (2,4- dioxies Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) butyramide (70)
According to 8 condition C of general step, false peptide 55 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 55 (34mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 70.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 99:1 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 70, (18mg, 19 μm ol, 46%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3293,2926,2853,1650,1554cm-1；1H NMR(400MHz,(CD3)2SO) δ, ca. rotation are different The 1:1 mixture of structure body) δ 11.29 (s, 1H, uracil-NH), 10.79 (s, 1H, Trp-NH), 8.56 (t, J=5.5Hz, 0.5H,CONHCH2), 8.24 (d, J=9.0Hz, 0.5H, CONH), 8.13-8.09 (m, 1H, 0.5x CONH+0.5x CONHCH2),8.05(br s,3H,NH3),7.75-7.72(m,1H,CONHCH2(CH2)4CH3),7.55-7.52(m,1.5H,1x H-6+0.5x Ar-H),7.52-7.50(m,0.5H,Ar-H),7.32-7.30(m,1H,Ar-H),7.08-7.02(m,2H,Ar- H),6.97-6.93(m,1H,Ar-H),6.35-6.33(m,1H,NHCONH),6.24-6.22(m,1H,NHCONH),5.71(d, J=3.7Hz, 0.5H, H-1 '), 5.67 (d, J=3.4Hz, 0.5H, H-1 '), 5.62-5.58 (m, 1H, H-5), 4.83-4.76 (m, 0.5H, DABA- β-CH), 4.59 (t, J=9.4Hz, 0.5H, DABA- α-CH), 4.51 (t, J=9.3Hz, 0.5H, DABA- α-CH),4.41-4.25(m,3H,1x Trp-α-CH+1x H-4’+1x H-2’),4.17-4.11(m,1H,Cha-α-CH), 3.90-3.77(m,2H,1.5x Gly-α-CH2+0.5x DABA-β-CH),3.58-3.55(m,0.5H,Gly-α-CH2), 3.30-3.24(m,1H,H-5’),3.18-3.12(m,1H,H-5’),3.03-2.88(m,4H,2x Trp-β-CH2+2x CONHCH2(CH2)4CH3),2.76(s,1.5H,NCH3),2.75(s,1.5H,NCH3),2.22-2.09(m,1H,H-3’), 1.69-1.60(m,6H,1x H-3’+5x Cy),1.44-1.29(m,2H,Cha-β-CH2),1.28-1.12(m,12.5H,3x Cy+1.5x CH3+8x CONHCH2(CH2)4CH3), 1.03 (d, J=6.9Hz, 1.5H, CH3),0.89-0.70(m,6H,3x Cy +3x CONHCH2(CH2)4CH3)；13C NMR(100MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.4, 172.3,170.3,169.9,166.5,166.1,163.7,163.7,158.9,158.6,158.3,157.6,157.6, 151.0,141.5,141.4,136.5,128.0,123.9,121.2,119.2,119.0,118.6,116.2,111.6, 110.5,102.2,101.9,92.5,92.3,79.1,74.5,74.4,55.2,54.8,54.5,53.2,51.7,51.6, 50.9,44.2,44.1,36.0,35.9,33.9,33.7,32.7,32.6,31.4,29.3,29.2,27.7,26.5,26.4, 26.3,26.1,22.5,15.0,14.4,14.2；LRMS[M+H]+865.8；HRMS C43H64N10O9Calculated value: MH+, 865.49305.Measured value: MH+,865.49207。
(2S, 3S) -2- ((S) -2- (3- ((S) -3- (1H- indol-3-yl) -1- (new penta) -1- oxo propyl- 2- yl) urea Base) -3- hexamethylene propionamide) -3- (2- Amino-N-methyl acetylamino)-N- (((2R, 4R, 5R) -5- (2,4- dioxos -3,4- Dihydro-pyrimidin -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) butyramide (71)
According to 8 condition C of general step, false peptide 56 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 56 (34mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 71.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 98:2 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 71, (20mg, 21 μm ol, 53%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3305,2925,1654,1551cm-1；1H NMR(400MHz,(CD3)2SO, ca. rotational isomer 1:1 mixture) δ 11.29 (s, 1H, uracil-NH), 10.80 (s, 1H, Trp-NH), 8.56 (t, J=5.5Hz, 0.5H, CONHCH2), 8.18 (d, J=9.3Hz, 0.5H, CONH), 8.12 (d, J=9.3Hz, 0.5H, CONH), 8.09-8.05 (m, 3.5H,3x NH3+0.5x CONHCH2),7.65-7.61(m,1H,CONHCH2C(CH3)3),7.57-7.50(m,2H,1x H-6+ 1x Ar-H),7.33-7.30(m,1H,Ar-H),7.11-7.10(m,1H,Ar-H),7.07-7.03(m,2H,Ar-H),6.98- 6.94 (m, 1H, Ar-H), 6.34-6.27 (m, 2H, NHCONH), 5.71 (d, J=3.7Hz, 0.5H, H-1 '), 5.67 (d, J= 3.5Hz, 0.5H, H-1 '), 5.62-5.58 (m, 1H, H-5), 4.82-4.75 (m, 0.5H, DABA- β-CH), 4.57 (t, J= 9.4Hz, 0.5H, DABA- α-CH), 4.49 (t, J=9.2Hz, 0.5H, DABA- α-CH), 4.44-4.38 (m, 1H, Trp- α- CH),4.36-4.24(m,2H,1x H-4’+1x H-2’),4.17-4.12(m,1H,Cha-α-CH),3.93-3.75(m,2H, 1.5x Gly-α-CH2+0.5x DABA-β-CH),3.59-3.53(m,0.5H,Gly-α-CH2),3.29-3.22(m,1H,H- 5 '), 3.18-3.11 (m, 1H, H-5 '), 3.02 (dd, J=14.6,6.3Hz, 1H, Trp- β-CH2),2.93-2.85(m,2H, 1x Trp-β-CH2+1x CONHCH2C(CH3)3), 2.80 (dd, J=13.2,6.3Hz, 1H, CONHCH2C(CH3)3),2.75 (s,1.5H,NCH3),2.74(s,1.5H,NCH3),2.22-2.10(m,1H,H-3’),1.69-1.60(m,6H,1x H-3’+5x Cy),1.43-1.30(m,2H,Cha-β-CH2),128-1.25(m,1H,1x Cy)1.16-1.10(m,3.5H,2x Cy+1.5x CH3), 1.00 (d, J=6.9Hz, 1.5H, CH3),0.92-0.82(m,2H,Cy),0.76(s,10H,9x CONHCH2C(CH3)3+ 1x Cy)；13C NMR(100MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.4,172.8,170.3, 169.9,166.5,166.1,163.7,163.7,159.0,158.7,158.4,158.1,157.7,157.6,151.0, 141.5,141.4,136.5,127.9,123.9,121.8,121.2,119.0,118.8,118.6,115.8,112.9, 111.6,110.6,102.2,101.9,92.5,92.3,79.1,74.5,74.4,55.2,54.9,54.5,53.2,51.6, 51.5,50.9,50.1,44.2,44.1,36.0,35.9,33.9,33.7,32.6,32.4,29.2,29.0,27.6,26.5, 26.3,26.1,15.0,14.2；LRMS[M+H]+851.8；HRMS C42H62N10O9Calculated value: MH+,851.47740.Measured value: MH+,851.7703。
(2S, 3S) -3- (2- Amino-N-methyl acetylamino) -2- ((S) -2- (3- ((S) -1- (benzylamino) -3- (1H- indol-3-yl) -1- oxo propyl- 2- yl) urea groups) -3- hexamethylene propionamide)-N- (((2R, 4R, 5R) -5- (2,4- dioxies Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) butyramide (72)
According to 8 condition C of general step, false peptide 57 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 57 (34mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 72.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 99:1 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 72, (13mg, 14 μm ol, 34%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3292,2924,1672,1550cm-1；1H NMR(400MHz,(CD3)2SO, ca. rotational isomer 1:1 mixture) δ 11.29 (s, 1H, uracil-NH), 10.82 (s, 1H, Trp-NH), 8.55 (t, J=5.3Hz, 0.5H, CONHCH2),8.34-8.31(m,1H,CONHCH2(C6H5)), 8.19 (d, J=9.2Hz, 0.5H, CONH), 8.14 (d, J= 9.2Hz,0.5H,CONH),8.09-8.03(m,3.5H,3x NH3+0.5CONHCH2),7.58-7.49(m,2H,1x Ar-H+1x H-6),7.35-7.30(m,1H,Ar-H),7.25-7.17(m,3H,Ar-H),7.08-7.03(m,4H,Ar-H),6.98-6.94 (m, 1H, Ar-H), 6.35-6.33 (m, 1H, NHCONH), 6.30-6.27 (m, 1H, NHCONH), 5.70 (d, J=3.4Hz, 0.5H, H-1 '), 5.66 (d, J=3.6Hz, 0.5H, H-1 '), 5.61-5.57 (m, 1H, H-5), 4.83-4.75 (m, 0.5H, DABA- β-CH), 4.58 (t, J=9.5Hz, 0.5H, DABA- α-CH), 4.50 (t, J=9.2Hz, 0.5H, DABA- α-CH), 4.44-4.36(m,1H,Trp-α-CH),4.32-4.26(m,2H,1x H-4’+1x H-2’),4.24-4.20(m,2H, CONHCH2(C6H5)),4.17-4.12(m,1H,Cha-α-CH),3.88-3.76(m,2H,1.5x Gly-α-CH2+0.5x DABA-β-CH),3.58-3.54(m,0.5H,Gly-α-CH2),3.29-3.22(m,1H,H-5’),3.17-3.11(m,1H,H- 5 '), 3.04 (dd, J=14.5,6.6Hz, 1H, Trp- β-CH2), 2.95 (dd, J=14.3,6.7Hz, 1H, Trp- β-CH2), 2.75(s,1.5H,NCH3),2.74(s,1.5H,NCH3),2.21-2.19(m,1H,H-3’),1.68-1.58(m,6H,1x H- 3’+5x Cy),1.44-1.29(m,2H,Cha-β-CH2),128-1.23(m,1H,1x Cy)1.16-1.09(m,3.5H,2x Cy+1.5x CH3), 1.02 (d, J=6.9Hz, 1.5H, CH3),0.88-0.76(m,3H,Cy)；13C NMR(100MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.4,172.7,170.3,169.9,166.5,166.1,163.7, 158.6,158.3,157.6,151.0,141.5,141.4,139.7,136.6,128.6,127.9,127.4,127.0, 124.1,121.3,119.1,118.7,116.3,111.7,110.4,102.2,101.9,92.5,92.3,79.1,74.5, 74.4,55.2,54.6,53.2,51.7,50.9,44.2,44.1,42.5,36.0,35.9,33.9,33.7,32.7,32.6, 29.3,27.7,26.5,26.3,26.1,15.0,14.2；LRMS[M+H]+871.8；HRMSC44H58N10O9Calculated value: MH+, 871.44610.Measured value: MH+,871.44632。
Methyl ((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- (((2R, 4R, 5R) -5- (2,4- Dioxo) -3,4- dihydro-pyrimidin -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo butyl- 2- yl) Amino) -3- (naphthalene -2- base) -1- oxopropan -2- base) carbamoyl)-L-Trp ester (73)
According to 8 condition C of general step, false peptide 58 is cracked from resin.Preactivated EDCHCl (13mg, 65 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (7.3 μ L, 65 μm of ol, 1.3eq.)2Cl2: DMF (160 μ L) solution is added dropwise to thick residual Excess 58 (42mg, 50 μm of ol, 1eq.), uridine amine 18 (34mg, 0.15mmol, 3eq.) and HOAt (34mg, 0.25mmol, 1:2v/vCH 5eq.)2Cl2: in DMF (160 μ L) solution, obtain shielded analog 73.Use 1:1v/vCH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 97:3 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 73, (13mg, 14 μm ol, 27%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3213,1673,1549,1440cm-1；1HNMR(500MHz,(CD3)2The 1 of SO, ca. rotational isomer: 1 mixture) δ 11.29 (s, 1H, uracil-NH), 10.89 (s, 1H, Trp-NH), 8.61 (t, J=5.1Hz, 0.5H, CONHCH2), 8.43 (d, J=9.1Hz, 0.5H, CONH), 8.38 (d, J=9.1Hz, 0.5H, CONH), 8.13 (t, J= 5.1Hz,,0.5H,CONHCH2),8.05(brs,3H,NH3),7.84-7.83(m,1H,Ar-H),7.77-7.75(m,2H,Ar- ), H 7.62 (m, 1H, Ar-H), 7.55 (d, J=8.1Hz, 0.5H, H-6), 7.51 (d, J=8.1Hz, 0.5H, H-6), 7.48- 7.42(m,3H,Ar-H),7.33-7.30(m,2H,Ar-H),7.11-7.04(m,2H,Ar-H),6.98-6.95(m,1H,Ar- ), H 6.56-6.54 (m, 1H, NHCONH), 6.43-6.39 (m, 1H, NHCONH), 5.72 (d, J=3.6Hz, 0.5H, H-1 '), 5.69 (d, J=3.4Hz, 0.5H, H-1 '), 5.60-5.57 (m, 1H, H-5), 4.81-4.78 (m, 0.5H, DABA- β-CH), 4.61 (t, J=9.4Hz, 0.5H, DABA- α-CH), 4.54-4.48 (m, 1.5H, 1xDABA- α-CH+1xTrp- α-CH), 4.41-4.33(m,2.5H,0.5xH-4’+1xH-2’+1xNal-α-CH),4.28-4.26(m,0.5H,H-4’),3.96-3.78 (m,2H,1.5xGly-α-CH2+0.5xDABA-β-CH),3.59-3.55(m,0.5H,Gly-α-CH2),3.52(s,1.5H, OCH3),3.51(s,1.5H,OCH3),3.26-3.12(m,2H,H-5’),3.11-3.07(m,1H,Trp-β-CH2),3.05- 2.98(m,2H,Nal-β-CH2),2.93-2.89(m,1H,Trp-β-CH2),2.78(s,1.5H,NCH3),2.75(s,1.5H, NCH3), 2.22-2.12 (m, 1H, H-3 '), and 1.70-1.62 (m, 1H, H-3 '), 1.14 (d, J=6.5Hz, 1.5H, CH3),1.04 (d, J=6.8Hz, 1.5H, CH3)；13CNMR(125MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.8, 172.1,170.3,170.0,166.7,166.3,163.9,163.8,158.7,158.5,157.5,151.2,141.8, 141.5,136.7,135.8,135.7,133.5,132.4,128.6,128.5,128.3,128.0,128.0,127.9, 127.9,127.8,126.4,125.9,124.4,121.5,119.0,118.7,112.0,109.7,102.4,102.0,92.6, 92.5,79.2,79.2,74.6,74.5,55.4,55.0,54.9,54.8,54.2,53.6,52.2,51.0,44.5,44.3, 38.9,38.7,36.3,36.1,29.4,28.4,27.9,15.2,14.4；LRMS[M+H]+840.4；HRMS calculated value C42H49N9O10: MH+,840.36752.Measured value: MH+,840.36862。
Hexyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- Base) amino) -3- (naphthalene -2- base) -1- oxopropan -2- base) carbamoyl)-L-Trp ester (74)
According to 8 condition C of general step, false peptide 59 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 59 (36mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/vCH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 74.Use 1:1v/vCH2Cl2: TFA (5.2mL) And 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable non-enantiomer mixture of 99:1.Pass through Reversed-phase HPLC purifying obtains white fluffy solid 74 (12mg, 12 μm of ol, 30%, HPLC production calculated after purification with tfa salt Rate).
IR(ATR):3350,2931,1679,1555cm-1；1HNMR(400MHz,(CD3)2The 1 of SO, ca. rotational isomer: 1 mixture) δ 11.29 (s, 1H, uracil-NH), 10.88 (s, 1H, Trp-NH), 8.61 (t, J=5.3Hz, 0.5H, CONHCH2), 8.44 (d, J=9.1Hz, 0.5H, CONH), 8.40 (d, J=9.2Hz, 0.5H, CONH), 8.13 (t, J= 5.6Hz,0.5H,CONHCH2),8.05(brs,3H,NH3),7.85-7.83(m,1H,Ar-H),7.78-7.75(m,2H,Ar- ), H 7.63 (m, 1H, Ar-H), 7.55 (d, J=8.1Hz, 0.5H, H-6), 7.52 (d, J=8.1Hz, 0.5H, H-6), 7.49- 7.42(m,3H,Ar-H),7.34-7.30(m,2H,Ar-H),7.11(m,1H,Ar-H),7.08-7.04(m,1H,Ar-H), 6.99-6.95 (m, 1H, Ar-H), 6.59-6.56 (m, 1H, NHCONH), 6.43-6.39 (m, 1H, NHCONH), 5.73 (d, J= 3.6Hz, 0.5H, H-1 '), 5.70 (d, J=3.5Hz, 0.5H, H-1 '), 5.61-5.58 (m, 1H, H-5), 4.84-4.77 (m, 0.5H, DABA- β-CH), 4.61 (t, J=9.4Hz, 0.5H, DABA- α-CH), 4.55-4.49 (m, 1.5H, 1xDABA- α-CH+ 1xTrp-α-CH),4.41-4.32(m,2.5H,0.5xH-4’+1xH-2’+1xNal-α-CH),4.29-4.25(m,0.5H,H- 4’),3.96-3.79(m,4H,1.5xGly-α-CH2+0.5xDABA-β-CH+2xOCH2(CH2)4CH3),3.64-3.51(m, 0.5H,Gly-α-CH2),3.27-3.12(m,2H,H-5’),3.10-3.06(m,1H,Trp-β-CH2),3.02-3.00(m,2H, Nal-β-CH2),2.97-2.90(m,1H,Trp-β-CH2),2.79(s,1.5H,NCH3),2.76(s,1.5H,NCH3),2.24- 2.12(m,1H,H-3’),1.71-1.62(m,1H,H-3’),1.39-1.37(m,2H,OCH2(CH2)4CH3),1.24-1.13 (m,7.5H,6xOCH2(CH2)4CH3+1.5xCH3), 1.04 (d, J=6.9Hz, 1.5H, CH3), 0.83 (t, J=6.8Hz, 3H, CH3)；13CNMR(100MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.3,171.9,170.2,169.8, 166.5,166.2,163.7,158.6,158.2,157.9,157.4,151.0,141.6,141.4,136.6,135.6, 135.5,133.4,132.3,128.4,128.1,127.9,127.8,127.8,127.7,127.6,126.2,125.8, 124.2,121.4,118.8,118.5,111.8,109.6,102.2,101.9,92.5,92.3,79.1,74.5,74.3, 64.7,55.3,54.9,54.7,54.6,54.1,53.4,50.9,44.3,44.2,38.8,38.6,36.1,35.9,31.3, 29.2,28.4,27.7,25.3,22.4,15.0,14.3；LRMS[M+H]+910.9；HRMSC47H59N9O10Calculated value: MH+, 910.44577.Measured value: MH+,910.44614。
Dodecyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) - 5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo fourth Alkane -2- base) amino) -3- (naphthalene -2- base) -1- oxopropan -2- base) carbamoyl)-L-Trp ester (75)
According to 8 condition C of general step, false peptide 60 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 60 (39mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/vCH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 75.Use 1:1v/vCH2Cl2: TFA (5.2mL) And 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable non-enantiomer mixture of 99:1.Pass through Reversed-phase HPLC purifying obtains white fluffy solid 75 (14mg, 13 μm of ol, 31%, HPLC production calculated after purification with tfa salt Rate).
IR(ATR):3305,2924,2854,1677,1557cm-1；1HNMR(500MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 11.29 (s, 1H, uracil-NH), 10.86 (s, 1H, Trp-NH), 8.60 (t, J=5.5Hz, 0.5H, CONHCH2), 8.44-8.39 (m, 1H, CONH), 8.13 (t, J=5.1Hz, 0.5H, CONHCH2),8.01(brs,3H,NH3), 7.84-7.82 (m, 1H, Ar-H), 7.77-7.74 (m, 2H, Ar-H), 7.62 (m, 1H, Ar-H), 7.54 (d, J=8.1Hz, 0.5H, H-6), 7.50 (d, J=8.1Hz, 0.5H, H-6), 7.48-7.42 (m, 3H, Ar-H), 7.33-7.29 (m, 2H, Ar- H),7.10(m,1H,Ar-H),7.06-7.03(m,1H,Ar-H),6.97-6.94(m,1H,Ar-H),6.58-6.55(m,1H, ), NHCONH 6.41-6.36 (m, 1H, NHCONH), 5.72 (d, J=3.6Hz, 0.5H, H-1 '), 5.69 (d, J=3.6Hz, 0.5H, H-1 '), 5.64-5.57 (m, 1H, H-5), 4.82-4.77 (m, 0.5H, DABA- β-CH), 4.61 (t, J=9.4Hz, 0.5H,DABA-α-CH),4.55-4.49(m,1.5H,0.5xDABA-α-CH+1xTrp-α-CH),4.41-4.33(m,2.5H, 0.5xH-4’+1xH-2’+1xNal-α-CH),4.30-4.25(m,0.5H,H-4’),3.95-3.78(m,4H,1.5xGly-α- CH2+0.5xDABA-β-CH+2xOCH2(CH2)10CH3),3.59-3.54(m,0.5H,Gly-α-CH2),3.27-3.12(m,2H, H-5’),3.10-3.05(m,1H,Trp-β-CH2),3.04-2.97(m,2H,Nal-β-CH2),2.94-2.90(m,1H,Trp- β-CH2),2.78(s,1.5H,NCH3),2.76(s,1.5H,NCH3),2.22-2.12(m,1H,H-3’),1.70-1.62(m, 1H,H-3’),1.41-1.36(m,2H,OCH2(CH2)10CH3),1.26-1.12(m,19.5H,18xOCH2(CH2)10CH3+ 1.5xCH3), 1.03 (d, J=6.9Hz, 1.5H, CH3), 0.84 (t, J=7.0Hz, 3H, CH3)；13CNMR(125MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.5,172.0,170.3,170.0,166.7,166.3,163.9, 163.8,158.8,158.5,158.3,157.5,151.2,141.7,141.5,136.7,135.7,135.6,133.5, 132.4,128.6,128.5,128.3,128.0,128.0,127.9,127.8,126.4,125.9,124.3,121.5, 118.9,118.7,112.0,109.7,102.4,102.0,92.6,92.5,79.2,74.6,74.5,64.8,55.4,55.0, 54.8,54.7,54.3,53.5,44.5,44.3,39.0,38.8,36.2,36.1,29.6,29.6,29.5,29.4,29.3, 29.2,28.5,27.9,25.8,22.7,15.2,14.6；LRMS[M+H]+995.1；HRMSC53H71N9O10Calculated value: MH+, 994.53967.Measured value: MH+,994.54051。
Neopentyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes - 2- yl) amino) -3- (naphthalene -2- base) -1- oxopropan -2- base) carbamoyl)-L-Trp ester (76)
According to 8 condition C of general step, false peptide 61 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 61 (35mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/vCH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 76.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 97:3 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 76, (13mg, 13 μm ol, 32%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3285,3063,2957,1676,1552cm-1；1HNMR(500MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 11.29 (s, 1H, uracil-NH), 10.86 (s, 1H, Trp-NH), 8.59 (t, J=5.5Hz, 0.5H, CONHCH2), 8.43 (d, J=9.0Hz, 0.5H, CONH), 8.39 (d, J=9.3Hz, 0.5H, CONH), 8.12 (t, J= 5.2Hz,0.5H,CONHCH2),8.02(brs,3H,NH3),7.84-7.82(m,1H,Ar-H),7.77-7.74(m,2H,Ar- ), H 7.62 (m, 1H, Ar-H), 7.54 (d, J=8.1Hz, 0.5H, H-6), 7.50 (d, J=8.1Hz, 0.5H, H-6), 7.46- 7.42(m,3H,Ar-H),7.33-7.29(m,2H,Ar-H),7.10(m,1H,Ar-H),7.07-7.04(m,1H,Ar-H), 6.98-6.95 (m, 1H, Ar-H), 6.57-6.54 (m, 1H, NHCONH), 6.43-6.39 (m, 1H, NHCONH), 5.72 (d, J= 3.7Hz, 0.5H, H-1 '), 5.69 (d, J=3.5Hz, 0.5H, H-1 '), 5.60-5.57 (m, 1H, H-5), 4.81-4.77 (m, 0.5H, DABA- β-CH), 4.60 (t, J=9.4Hz, 0.5H, DABA- α-CH), 4.55-4.49 (m, 1.5H, 0.5xDABA- α- CH+1xTrp-α-CH),4.45-4.41(m,1H,Nal-α-CH),4.38-4.31(m,1.5H,0.5xH-4’+1xH-2’), 4.29-4.24(m,0.5H,H-4’),3.96-3.77(m,2H,1.5xGly-α-CH2+0.5xDABA-β-CH),3.61-3.57 (m,2.5H,2xOCH2C(CH3)3+0.5xGly-α-CH2),3.26-3.11(m,2H,H-5’),3.09-3.01(m,3H, 1xTrp-β-CH2+2xNal-β-CH2),2.94-2.89(m,1H,Trp-β-CH2),2.78(s,1.5H,NCH3),2.75(s, 1.5H,NCH3), 2.22-2.12 (m, 1H, H-3 '), 1.70-1.62 (m, 1H, H-3 '), 1.12 (d, J=6.6Hz, 1.5H, CH3), 1.03 (d, J=6.9Hz, 1.5H, CH3),0.75(s,4.5H,OCH2C(CH3)3),0.74(s,4.5H,OCH2C (CH3)3)；13CNMR(125MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 173.4,172.1,170.3, 170.0,166.7,166.3,163.9,163.8,158.7,158.4,157.5,151.2,141.7,141.5,136.8, 135.7,135.6,133.5,132.5,128.5,128.5,128.2,128.0,127.9,127.8,126.4,125.9, 124.3,121.5,119.0,118.7,112.0,109.8,102.4,102.0,92.6,92.5,79.2,74.6,74.5, 73.8,55.4,55.0,54.8,54.7,54.3,53.5,51.0,44.5,44.3,39.0,38.8,36.2,36.1,31.5, 29.4,28.6,27.9,26.6,15.2,14.4；LRMS[M+H]+896.8；HRMS calculated value C46H57N9O10: MH+, 896.43012.Measured value: MH+,896.43189。
Benzyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- Base) amino) -3- (naphthalene -2- base) -1- oxopropan -2- base) carbamoyl)-L-Trp ester (77)
According to 8 condition C of general step, false peptide 62 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 62 (36mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 77.Use 1:1v/vCH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 99:1 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 77, (11mg, 11 μm ol, 27%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3331,3057,2945,1677,1550cm-1；1HNMR(500MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 11.29 (s, 1H, uracil-NH), 10.89 (s, 1H, Trp-NH), 8.61 (t, J=5.0Hz, 0.5H, CONHCH2), 8.44 (d, J=9.0Hz, 0.5H, CONH), 8.41 (d, J=9.2Hz, 0.5H, CONH), 8.13 (t, J= 5.6Hz,0.5H,CONHCH2),8.03(brs,3H,NH3),7.83-7.82(m,1H,Ar-H),7.75-7.72(m,2H,Ar- ), H 7.60 (m, 1H, Ar-H), 7.55 (d, J=8.0Hz, 0.5H, H-6), 7.51 (d, J=8.1Hz, 0.5H, H-6), 7.47- 7.42(m,3H,Ar-H),7.35-7.34(m,1H,Ar-H),7.30-7.23(m,4H,Ar-H),7.16-15(m,2H,Ar-H), 7.08-7.05(m,2H,Ar-H),6.98-6.95(m,1H,Ar-H),6.64--6.61(m,1H,NHCONH),6.42-6.38 (m, 1H, NHCONH), 5.72 (d, J=3.5Hz, 0.5H, H-1 '), 5.69 (d, J=3.3Hz, 0.5H, H-1 '), 5.60-5.57 (m,1H,H-5),5.02-4.95(m,2H,OCH2(C6H5)), 4.83-4.79 (m, 0.5H, DABA- β-CH), 4.61 (t, J= 9.3Hz,0.5H,DABA-α-CH),4.56-4.50(m,1.5H,0.5xDABA-α-CH+1xTrp-α-CH),4.48-4.44(m, 1H,Nal-α-CH),4.39-4.33(m,1.5H,0.5xH-4’+1xH-2’),4.30-4.25(m,0.5H,H-4’),3.96- 3.78(m,2H,1.5xGly-α-CH2+0.5xDABA-β-CH),3.60-3.55(m,0.5H,Gly-α-CH2),3.25-3.11 (m,2H,H-5’),3.10-3.01(m,3H,1xTrp-β-CH2+2xNal-β-CH2),2.94-2.90(m,1H,Trp-β-CH2), 2.78(s,1.5H,NCH3),2.75(s,1.5H,NCH3),2.22-2.12(m,1H,H-3’),1.70-1.62(m,1H,H-3’), 1.13 (d, J=6.4Hz, 1.5H, CH3), 1.03 (d, J=6.8Hz, 1.5H, CH3)；13CNMR(125MHz,(CD3)2SO,ca. The 1:1 mixture of rotational isomer) δ 173.3,172.0,170.3,170.0,166.7,166.3,163.9,163.8,158.9, 158.7,158.4,157.5,151.2,141.7,141.5,136.8,136.4,135.7,135.6,133.5,132.4, 129.5,128.9,128.8,128.5,128.3,128.3,128.0,128.0,127.9,127.9,127.8,126.4, 125.9,124.5,121.6,119.1,118.7,112.0,109.7,102.4,102.0,92.6,92.5,79.2,74.6, 74.5,67.6,66.4,55.4,55.0,54.8,54.7,54.4,53.6,51.0,44.5,44.3,39.0,38.8,36.2, 36.1,31.3,29.4,28.5,27.9,25.7,15.2,14.5；LRMS[M+H]+916.8；HRMS calculated value C48H53N9O10: MH+,916.39882.Measured value: MH+,916.39964。
(2S, 3S) -3- (2- Amino-N-methyl acetylamino)-N- (((2R, 4R, 5R) -5- (2,4- dioxos -3,4- bis- Hydrogen pyrimidine -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) -2- ((S) -2- (3- ((S) -1- (hexylamino) -3- (1H- indol-3-yl) -1- oxo propyl- 2- yl) urea groups) -3- (naphthalene -2- base) propionamido) butyramide (78)
According to 8 condition C of general step, false peptide 63 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 63 (36mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/v CH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 78.Use 1:1v/vCH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 98:2 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 78, (22mg, 22 μm ol, 54%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3292,2930,1663,1551cm-1；1HNMR(400MHz,(CD3)2The 1 of SO, ca. rotational isomer: 1 mixture) δ 11.29 (s, 1H, uracil-NH), 10.78 (s, 1H, Trp-NH), 8.59 (t, J=5.2Hz, 0.5H, CONHCH2), 8.37 (d, J=9.1Hz, 0.5H, CONH), 8.33 (d, J=9.1Hz, 0.5H, CONH), 8.10 (t, J= 5.2Hz,,0.5H,CONHCH2),8.05(brs,3H,NH3), 7.84-7.74 (m, 3H, Ar-H), 7.68 (t, J=5.5Hz, 1H, CONHCH2(CH2)4CH3),7.63(m,1H,Ar-H),7.55-7.51(m,2H,1xH-6+1xAr-H),7.49-7.42(m,2H, Ar-H),7.33-7.29(m,2H,Ar-H),7.06-7.01(m,2H,Ar-H),6.96-6.92(m,1H,Ar-H),6.42- 6.40 (m, 2H, NHCONH), 5.72 (d, J=3.7Hz, 0.5H, H-1 '), 5.69 (d, J=3.5Hz, 0.5H, H-1 '), 5.61- 5.57 (m, 1H, H-5), 4.82-4.73 (m, 0.5H, DABA- β-CH), 4.61 (t, J=9.3Hz, 0.5H, DABA- α-CH), 4.55-4.41(m,1.5H,1xDABA-α-CH+1xTrp-α-CH),4.38-4.24(m,3H,1xH-4’+1xH-2’+1xNal- α-CH),3.91-3.77(m,2H,1.5xGly-α-CH2+0.5xDABA-β-CH),3.59-3.54(m,0.5H,Gly-α-CH2), 3.27-3.07(m,3H,2xH-5’+1xCONHCH2(CH2)4CH3),3.00-2.85(m,5H,2xTrp-β-CH2+2xNal-β- CH2+1xCONHCH2(CH2)4CH3),2.77(s,1.5H,NCH3),2.76(s,1.5H,NCH3),2.22-2.11(m,1H,H- 3’),1.70-1.61(m,1H,H-3’),1.24-1.13(m,9.5H,8xCONHCH2(CH2)4CH3+1.5xCH3),1.03(d,J =6.8Hz, 1.5H, CH3), 0.81 (t, J=6.9Hz, 3H, CONHCH2(CH2)4CH3)；13CNMR(100MHz,(CD3)2SO, Ca. the 1:1 mixture of rotational isomer) δ 172.3,172.1,170.2,169.8,166.5,166.1,163.7,163.7, 159.1,158.8,158.6,158.5,158.1,157.4,151.0,141.6,141.4,136.5,135.8,135.7, 135.0,133.4,132.3,128.4,128.1,127.9,127.9,127.8,127.8,126.3,125.8,123.9, 121.2,119.0,118.7,118.5,115.7,113.9,111.6,110.5,102.2,101.9,92.5,92.3,79.1, 79.0,74.5,74.3,63.0,55.5,55.3,54.9,54.8,54.5,53.4,50.9,44.3,44.1,38.9,38.7, 36.1,35.9,31.4,29.3,29.2,27.7,26.4,22.4,15.0,14.4,14.2；LRMS[M+H]+909.9；HRMS meter Calculation value C47H60N10O9: MH+,909.47175.Measured value: MH+,909.46152。
(2S, 3S) -2- ((S) -2- (3- ((S) -3- (1H- indol-3-yl) -1- (new penta) -1- oxo propyl- 2- yl) urea Base) -3- (naphthalene -2- base) propionamido) -3- (2- Amino-N-methyl acetylamino)-N- (((2R, 4R, 5R) -5- (2,4- dioxies Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) butyramide (79)
According to 8 condition C of general step, false peptide 64 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 64 (35mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/vCH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 79.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 98:2 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 79, (13mg, 13 μm ol, 32%, HPLC is calculated after purification with tfa salt Yield).
IR(ATR):3292,2935,1672,1556cm-1；1HNMR(400MHz,(CD3)2The 1 of SO, ca. rotational isomer: 1 mixture) δ 11.31 (s, 1H, uracil-NH), 10.79 (s, 1H, Trp-NH), 8.60 (t, J=5.6Hz, 0.5H, CONHCH2), 8.40 (d, J=8.9Hz, 0.5H, CONH), 8.34 (d, J=9.1Hz, 0.5H, CONH), 8.13 (t, J= 5.6Hz,0.5H,CONHCH2),8.03(brs,3H,NH3),7.85-7.83(m,1H,Ar-H),7.79-7.74(m,2H,Ar- H),7.64-7.61(m,2H,1xAr-H+1xCONHCH2C(CH3)3),7.57-7.54(m,1.5H,0.5xH-6+1xAr-H), 7.51 (d, J=8.1Hz, 0.5H, H-6), 7.49-7.42 (m, 2H, Ar-H), 7.33-7.30 (m, 2H, Ar-H), 7.10-7.09 (m,1H,Ar-H),7.06-7.02(m,1H,Ar-H),6.97-6.94(m,1H,Ar-H),6.47--6.44(m,1H, ), NHCONH 6.40-6.37 (m, 1H, NHCONH), 5.72 (d, J=3.6Hz, 0.5H, H-1 '), 5.69 (d, J=3.5Hz, 0.5H, H-1 '), 5.62-5.58 (m, 1H, H-5), 4.84-4.76 (m, 0.5H, DABA- β-CH), 4.61 (t, J=9.4Hz, 0.5H,DABA-α-CH),4.55-4.48(m,1.5H,0.5xDABA-α-CH+1xTrp-α-CH),4.43-4.38(m,1H, Nal-α-CH),4.37-4.31(m,1.5H,0.5xH-4’+1xH-2’),4.29-4.24(m,0.5H,H-4’),3.92-3.77 (m,2H,1.5xGly-α-CH2+0.5xDABA-β-CH),3.59-3.54(m,0.5H,Gly-α-CH2),3.28-3.14(m,2H, H-5’),3.11-3.06(m,1H,Trp-β-CH2),3.03-2.98(m,1H,Nal-β-CH2),2.94-2.85(m,2H, 1xTrp-β-CH2+1xNal-β-CH2),2.83-2.78(m,2H,CONHCH2C(CH3)3),2.77(s,1.5H,NCH3),2.76 (s,1.5H,NCH3), 2.23-2.11 (m, 1H, H-3 '), 1.70-1.62 (m, 1H, H-3 '), 1.13 (d, J=6.5Hz, 1.5H, CH3), 1.02 (d, J=6.9Hz, 1.5H, CH3),0.72(s,4.5H,CONHCH2C(CH3)3),0.71(s,4.5H,CONHCH2C (CH3)3)；13CNMR(100MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 172.8,172.0,170.2, 169.8,166.5,166.1,163.7,159.0,158.7,158.3,158.0,157.5,151.0,141.6,141.4, 136.5,135.7,135.6,133.4,132.3,128.4,128.1,127.9,127.8,127.8,127.7,126.2, 125.8,123.9,121.2,119.0,118.7,118.6,115.7,111.6,110.6,102.2,101.9,92.4,92.3, 79.0,79.0,74.5,74.3,55.3,54.8,54.7,54.4,53.4,50.9,50.1,44.3,44.1,38.8,38.5, 36.1,35.9,32.3,29.2,29.1,27.7,27.6,15.0,14.2；LRMS[M+H]+895.8；HRMSC46H58N10O9It calculates Value: MH+,895.44610.Measured value: MH+,895.44543。
(2S, 3S) -3- (2- Amino-N-methyl acetylamino) -2- ((S) -2- (3- ((S) -1- (benzylamino) -3- (1H- indol-3-yl) -1- oxo propyl- 2- yl) urea groups) -3- (naphthalene -2- base) propionamido-)-N- (((2R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) butyramide (80)
According to 8 condition C of general step, false peptide 65 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 65 (36mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/vCH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 80.Use 1:1v/vCH2Cl2: TFA (5.2mL) And 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable non-enantiomer mixture of 97:3.Pass through Reversed-phase HPLC purifying obtains white fluffy solid 80 (12mg, 12 μm of ol, 29%, HPLC production calculated after purification with tfa salt Rate).
IR(ATR):3293,3041,2937,1655,1549cm-1；1HNMR(500MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 11.29 (s, 1H, uracil-NH), 10.81 (s, 1H, Trp-NH), 8.59 (t, J=5.5Hz, 0.5H, CONHCH2),8.38-8.28(m,2H,1xCONH+1xCONHCH2(C6H5)), 8.10 (t, J=5.5Hz, 0.5H, CONHCH2), 8.03(brs,3H,NH3),7.84-7.82(m,1H,Ar-H),7.77-7.73(m,2H,Ar-H),7.62(m,1H,Ar-H), 7.58-7.56 (m, 1H, Ar-H), 7.54 (d, J=8.1Hz, 0.5H, H-6), 7.50 (d, J=8.1Hz, 0.5H, H-6), 7.47-7.42(m,2H,Ar-H),7.34-7.30(m,2H,Ar-H),7.22-7.14(m,3H,Ar-H),7.07-7.03(m, 4H,Ar-H),6.97-6.93(m,1H,Ar-H),6.48-6.45(m,1H,NHCONH),6.40-6.37(m,1H,NHCONH), 5.71 (d, J=3.7Hz, 0.5H, H-1 '), 5.69 (d, J=3.5Hz, 0.5H, H-1 '), 5.61-5.57 (m, 1H, H-5), 4.84-4.76 (m, 0.5H, DABA- β-CH), 4.61 (t, J=9.3Hz, 0.5H, DABA- α-CH), 4.55-4.46 (m, 1.5H, 0.5xDABA-α-CH+1xTrp-α-CH),4.44-4.38(m,1H,Nal-α-CH),4.37-4.30(m,1.5H,0.5xH-4’+ 1xH-2’),4.28-4.24(m,0.5H,H-4’),4.21-4.14(m,2H,CONHCH2(C6H5)),3.95-3.76(m,2H, 1.5xGly-α-CH2+0.5xDABA-β-CH),3.57-3.54(m,0.5H,Gly-α-CH2),3.26-3.14(m,2H,H-5’), 3.12-3.07(m,1H,Trp-β-CH2),3.06-3.01(m,1H,Nal-β-CH2),2.95-2.90(m,2H,1xTrp-β-CH2 +1xNal-β-CH2),2.77(s,1.5H,NCH3),2.75(s,1.5H,NCH3),2.23-2.11(m,1H,H-3’),1.69- 1.61 (m, 1H, H-3 '), 1.13 (d, J=6.5Hz, 1.5H, CH3), 1.03 (d, J=6.9Hz, 1.5H, CH3)；13CNMR (125MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 172.7,172.0,170.2,166.1,163.7, 163.7,158.5,158.2,157.9,157.5,151.0,141.6,139.7,136.5,135.7,133.4,132.3, 128.6,128.1,127.9,127.8,127.4,127.0,126.3,125.8,124.1,121.3,119.1,118.6, 111.7,110.4,102.2,101.9,92.5,92.3,79.0,74.5,74.3,55.3,54.9,54.9,54.6,53.4, 50.9,44.3,44.1,42.4,38.7,38.5,36.1,35.9,29.2,27.7,15.0,14.2；LRMS[M+H]+915.8； HRMS C48H54N10O9Calculated value: MH+,915.41480.Measured value: MH+,915.41373。
Benzyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- (((2R, 4R, 5R) -5- (2, 4- dioxo) -3,4- dihydro-pyrimidin -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo butyl- 2- Base) amino) -3- (6,7- dimethoxy -2- oxo -2H- chromene -4- base) -1- oxopropan -2- base) carbamoyl)-L- Tryptophan ester (81)
According to 8 condition C of general step, false peptide 50 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 50 (39mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/vCH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 81.With 1:1v/v CH2Cl2: TFA (5.2mL) and 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable diastereoisomer mixing of 95:5 Object.It is purified by reversed-phase HPLC, obtaining white fluffy solid 81, (9.2mg, 8.3 μm ol, 21%, HPLC after purification in terms of tfa salt The yield of calculation).
IR(ATR):3332,2929,1676,1556cm-1；1HNMR(500MHz,(CD3)2The 1 of SO, ca. rotational isomer: 1 mixture) δ 11.29 (s, 1H, uracil-NH), 10.88 (s, 1H, Trp-NH), 8.57 (t, J=5.7Hz, 0.5H, CONHCH2), 8.42-8.38 (m, 2H, CONH), 8.12 (t, J=5.8Hz, 0.5H, CONHCH2),7.99-7.97(m,3H, NH3), 7.53 (d, J=8.1Hz, 0.5H, H-6), 7.49 (d, J=8.1Hz, 0.5H, H-6), 7.45-7.42 (m, 2H, Ar- H),7.35-7.34(m,1H,Ar-H),7.26-7.24(m,3H,Ar-H),7.16-7.13(m,2H,Ar-H),7.08-7.05 (m,2H,Ar-H),7.01(m,1H,Ar-H),6.98-6.95(m,1H,Ar-H),6.70-6.65(m,2H,NHCONH),5.95 (s, 0.5H, H-3 "), 5.94 (s, 0.5H, H-3 "), 5.69 (d, J=3.6Hz, 0.5H, H-1 '), 5.65 (d, J=3.4Hz, 0.5H,H-1’),5.61-5.57(m,1H,H-5),4.99-4.98(m,2H,OCH2(C6H5)),4.70-4.67(m,0.5H, DABA-β-CH),4.56-4.47(m,3H,1xDABA-α-CH+1xTrp-α-CH+1xH-10”),4.37-4.28(m,1.5H, 0.5xH-4’+1xH-2’),4.22-4.17(m,0.5H,H-4’),3.84-3.73(m,8.5H,2xGly-α-CH2+ 0.5xDABA-β-CH+6xOCH3),3.33-3.21(m,1H,H-5’),3.13-2.99(m,4H,2xTrp-β-CH2+1xH-5’+ 1xH-9”),2.94-2.86(m,1H,H-9”),2.74(s,1.5H,NCH3),2.72(s,1.5H,NCH3),2.22-2.17(m, 0.5H, H-3 '), 2.15-2.10 (m, 0.5H, H-3 '), 1.67-1.59 (m, 1H, H-3 '), 1.08 (d, J=6.5Hz, 1.5H, CH3), 0.98 (d, J=6.9Hz, 1.5H, CH3)；13CNMR(125MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ173.1,171.0,170.8,170.0,169.6,166.6,166.6,162.2,164.0,163.9,160.9,160.8, 158.8,158.6,157.6,153.0,152.6,151.2,149.6,146.5,141.7,136.8,136.3,128.9, 128.6,128.3,128.3,127.8,124.4,121.6,119.1,118.7,112.7,112.1,111.9,109.5, 106.9,102.4,102.0,100.7,92.5,79.3,79.2,74.7,74.5,66.5,56.7,56.5,55.4,54.9, 54.5,53.5,44.3,36.2,35.9,31.3,28.5,27.9,15.1,14.4；LRMS[M+H]+994.9；HRMS calculated value C49H55N9O14: MH+,994.39412.Measured value: MH+,994.39394。
Resin-bonded false peptide (82)
According to 7.2 condition A of general step, the false peptide 48 of resin-bonded is existed with DIC (150 μ L, 0.94mmol, 5eq.) CH2Cl2Pre-activate in (1.9mL), be then added neopentyl alcohol (83 μ L, 0.94mmol, 5eq.) and DMAP (2.3mg, 19mol, CH 0.1eq.)2Cl2(1.9mL) solution obtains the false peptide 82 of resin-bonded.
Neopentyl (((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- (((2R, 4R, 5R) -5- (2,4- dioxo) -3,4- dihydro-pyrimidin -1 (2H)-yl) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo butyl- 2- yl) amino) -3- cyclohexyl -1- oxo propyl- 2- base carbamoyl)-L-Trp salt (83)
According to 8 condition C of general step, false peptide 82 is cracked from resin.Preactivated EDCHCl (10mg, 52 μm of ol, 1.3eq.) and the 2:1v/v CH of NMM (5.7 μ L, 52 μm of ol, 1.3eq.)2Cl2: DMF (210 μ L) solution is added dropwise to thick residual Excess 82 (34mg, 40 μm of ol, 1eq.), uridine amine 18 (27mg, 120 μm of ol, 3eq.) and HOAt (27mg, 200 μm of ol, 5eq.) 1:2v/vCH2Cl2: in DMF (210 μ L) solution, obtain shielded analog 83.Use 1:1v/vCH2Cl2: TFA (5.2mL) And 2.5vol.%iPr3SiH (0.13mL) processing deprotection obtains the separable non-enantiomer mixture of 96:4.Pass through Reversed-phase HPLC purifying obtains white fluffy solid 83 (15mg, 16 μm of ol, 39%, HPLC production calculated after purification with tfa salt Rate).
IR(ATR):3293,2953,2852,1668,1557cm-1；1HNMR(400MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 11.29 (s, 1H, uracil-NH), 10.89 (s, 1H, Trp-NH), 8.54 (t, J=5.6Hz, 0.5H, CONHCH2), 8.22 (d, J=8.9Hz, 0.5H, CONH), 8.17 (d, J=9.2Hz, 0.5H, CONH), 8.03 (brs, 3.5H, 0.5xCONHCH2+3xNH3),7.54-7.50(m,1H,H-6),7.46-7.44(m,1H,Ar-H),7.34-7.32(m,1H,Ar- ), H 7.12-7.11 (m, 1H, Ar-H), 7.07-7.04 (m, 1H, Ar-H), 6.99-6.96 (m, 1H, Ar-H), 6.38 (d, J= 7.9Hz, 1H, NHCONH), 6.35-6.31 (m, 1H, NHCONH), 5.70 (d, J=3.5Hz, 0.5H, H-1 '), 5.66 (d, J= 3.5Hz, 0.5H, H-1 '), 5.61-5.58 (m, 1H, H-5), 4.82-4.75 (m, 0.5H, DABA- β-CH), 4.56 (t, J= 9.4Hz,0.5H,DABA-α-CH),4.45-4.41(m,1.5H,0.5xDABA-α-CH+1xTrp-α-CH),4.39-4.24(m, 2H,1xH-4’+1xH-2’),4.14-4.10(m,1H,Cha-α-CH),3.94-3.76(m,2H,1.5xGly-α-CH2+ 0.5xDABA-β-CH),3.66-3.54(m,2.5H,0.5xGly-α-CH2+2xOCH2C(CH3)3),3.29-3.21(m,1H,H- 5’),3.18-3.11(m,1H,H-5’),3.09-3.01(m,2H,Trp-β-CH2),2.76(s,1.5H,NCH3),2.74(s, 1.5H,NCH3),2.21-2.11(m,1H,H-3’),1.68-1.59(m,6H,1xH-3’+5xCy),1.37-1.28(m,2H, Cha-β-CH2),1.26-1.23(m,1H,Cy),1.14-1.10(m,3.5H,1.5xCH3+ 2xCy), 1.01 (d, J=6.8Hz, 1.5H,CH3),0.87-0.80(m,12H,3xCy+9xOCH2C(CH3)3)；13CNMR(100MHz,(CD3)2SO, ca. rotational isomeric The 1:1 mixture of body) δ 173.4,173.3,170.3,169.9,166.5,166.2,163.7,163.7,158.9,15 8.6, 158.3,158.0,157.6,157.6,151.0,141.6,141.4,136.6,127.6,124.2,121.4,118.8, 118.5,111.9,109.6,102.2,101.9,92.5,92.3,79.1,74.5,74.4,73.7,55.2,54.9,54.2, 53.2,51.5,50.8,44.2,44.1,36.0,35.9,33.8,33.7,33.7,32.7,32.6,31.4,29.2,28.4, 27.7,26.5,26.3,26.1,15.0,14.3；LRMS[M+H]+852.8；HRMS calculated value C42H61N9O10: MH+, 852.46142.Measured value: MH+,852.46191。
General step 8
Condition D:
By the peptide of resin-bonded 95:2.5:2.5v/v/v TFA:H2O:TIS concussion processing 1 hour.Then use TFA: H2O:TIS (× 10) washs resin, merges with cracked solution and is concentrated.The thick residue as obtained by reversed-phase HPLC purifying.
(((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- base) ammonia Base) -3- cyclohexyl -1- oxopropan -2- base) carbamoyl)-L-Trp (99)
According to 2 condition B of general step, using PyBOP (110mg, 0.21mmol, 4eq.) and NMM (23 μ L, 0.21mmol, Connexon 88 (50mg, 0.21mmol, 4eq.) 4eq.) is connected to rink amide resin (112mg, 0.053mmol, 1eq.) On, then in 1,2- dichloroethanes (0.60mL), PPTS (15mg, 0.058mmol, 1.1eq.) is loaded to 87 (60mg, 0.13mmol, 2.5eq.).Fmoc deprotection after, according to step 6 condition A, in DMF (0.40mL), using HATU (15mg, 0.040mmol, 2eq.) and iPr2EtN (11 μ L, 0.060mmol, 3eq.) and 20 (18mg, 0.040mmol, 2eq.) is coupled.So Afterwards according to general step 5, containing PhSiH3The CH of (49 μ L, 0.40mmol, 20eq.)2Cl2In (0.50mL), resin Pd (PPh3)4(4.6mg, 4.0 μm of ol, 0.2eq.) processing in DMF (0.4mL), is used then according to 6 condition A of general step HATU (15mg, 0.040mmol, 2eq.) and iPr2EtN and (11 μ L, 0.060mmol, 8eq.) and Boc-Gly-OH (7.0mg, 0.040mmol, 2eq.) coupling.Resin is subjected to Fmoc deprotection, is made in DMF (0.8mL) according to 6 condition B of general step With PyBOP (42mg, 0.08mmol, 4eq.) and NMM (18 μ L, 0.16mmol, 8eq.) and Fmoc-Cha-OH (32mg, 0.080mmol, 4eq.) coupling.After Fmoc deprotection, iPr is used in DMF (0.40mL) according to general step 42EtN(14 μ L, 0.080mmol, 4eq.) realize carbamate S12 (21mg, 0.040mmol, 2eq.) coupling.Then according to general step Described in rapid 8 condition E, the peptide of resin-bonded is cracked from resin, obtains crude residue, it is pure by reversed-phase HPLC Change, obtains fluffy white solid 99 (8.8mg, 11 μm of ol, 53%, yield is calculated with FA salt after purification in HPLC).
1H NMR(400MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 11.29 (br s, 1H, uracil- ), NH 10.89 (s, 1H, Trp-NH), 8.57 (t, J=5.5Hz, 0.5H, CONHCH2), 8.21 (d, J=9.0Hz, 0.5H, ), CONH 8.15 (d, J=9.1Hz, 0.5H, CONH), 8.08 (t, J=5.4Hz, 0.5H, CONHCH2), 7.54 (d, J= 8.1Hz,1H,H-6),7.52-7.50(m,1H,Ar-H),7.34-7.32(m,1H,Ar-H),7.12-7.11(m,1H,Ar-H), 7.07-7.03 (m, 1H, Ar-H), 6.98-6.94 (m, 1H, Ar-H), 6.38 (d, J=7.9Hz, 1H, NHCONH), 6.24- 6.21 (m, 1H, NHCONH), 5.70 (d, J=3.7Hz, 0.5H, H-1 '), 5.66 (d, J=3.4Hz, 0.5H, H-1 '), 5.61 (d, J=8.0Hz, 0.5H, H-5), 5.58 (d, J=8.0Hz, 0.5H, H-5), 4.82-4.74 (m, 0.5H, DABA- β-CH), 4.57 (t, J=9.4Hz, 0.5H, DABA- α-CH), 4.49 (t, J=9.3Hz, 0.5H, DABA- α-CH), 4.40-4.24 (m, 3H,1x Trp-α-CH+1x H-2’+1x H-4’),4.17-4.11(m,1H,Cha-α-CH),3.92-3.84(m,1.5H,1x Gly-α-CH2+ 0.5x DABA- β-CH), 3.76 (d, J=16.4Hz, 0.5H, Gly- α-CH2), 3.55 (d, J=16.0Hz, 0.5H,Gly-α-CH2),3.26-3.25(m,1H,H-5’),3.18-3.00(m,3H,1x H-5’+2x Trp-β-CH2),2.75 (s,1.5H,NCH3),2.74(s,1.5H,NCH3),2.22-2.10(m,1H,H-3’),1.69-1.59(m,6H,1x H-3’+5x Cy),1.41-1.27(m,3H,1x Cy+2x Cha-β-CH2),1.16-1.01(m,3.5H,2x Cy+1.5CH3),1.02(d,J =6.9Hz, 1.5H, CH3),0.88-0.76(m,3H,3x Cy)；13C NMR(100MHz,(CD3)2SO, ca. rotational isomer 1:1 mixture) δ 174.1,172.9,169.8,169.4,166.0,165.7,163.3,163.2,157.7,15 7.2,157.1, 150.5,141.1,141.0,136.0,127.4,127.3,123.8,120.8,118.4,118.3,111.3,109.4, 101.7,101.4,91.9,91.8,78.6,74.0,73.9,54.8,54.4,53.4,52.8,51.1,50.4,43.8,43.6, 41.4,35.6,35.4,33.4,33.2,32.1,28.7,27.2,26.0,25.8,25.7,22.9,14.5,13.8；LRMS[M+ H]+782.0..These data and Tran et al.[4]The data of report are consistent.
(((S) -1- (((2S, 3S) -3- (2- Amino-N-methyl acetylamino) -1- ((((2R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) -4- hydroxyl tetrahydrofuran -2- base) methyl) amino) -1- oxo-butanes -2- base) ammonia Base) -3- (naphthalene -2- base) -1- oxopropan -2- base) carbamoyl)-L-Trp (100)
According to 2 condition B of general step, using PyBOP (32mg, 0.062mmol, 4eq.) and NMM (7 μ L, 0.062mmol, Connexon 88 (15mg, 0.062mmol, 4eq.) 4eq.) is connected to rink amide resin (30mg, 0.016mmol, 1eq.) On, then in 1,2- dichloroethanes (0.12mL), PPTS (3mg, 0.012mmol, 1.1eq.) is loaded to 87 (17mg, 0.038mmol,2.5eq.).Fmoc deprotection after, according to step 6 condition A, in DMF (0.20mL), using HATU (8mg, 0.021mmol, 2eq.) and iPr2EtN (5.5 μ L, 0.032mmol, 3eq.) and 20 (9.2mg, 0.021mmol, 2eq.) is coupled. Then according to general step 5, containing PhSiH3The CH of (26 μ L, 0.21mmol, 20eq.)2Cl2In (0.26mL), resin Pd (PPh3)4(2.4mg, 2.1 μm of ol, 0.2eq.) processing in DMF (0.20mL), is used then according to 6 condition A of general step HATU (8mg, 0.021mmol, 2eq.) and iPr2EtN (5.5 μ L, 0.032mmol, 3eq.) and Boc-Gly-OH (3.7mg, 0.021mmol, 2eq.) coupling.According to 6 condition B of general step, resin is subjected to Fmoc deprotection, in DMF (0.40mL), Using PyBOP (22mg, 0.042mmol, 4eq.) and NMM (9 μ L, 0.084mmol, 8eq..) and Fmoc-Cha-OH (19mg, 0.042mmol, 4eq.) coupling.After Fmoc deprotection, iPr is used in DMF (0.20mL) according to general step 42EtN(7μ L, 0.042mmol, 4eq.) realize carbamate S12 (11mg, 0.021mmol, 2eq.) coupling.Then according to general step Described in rapid 8 condition E, the peptide of resin-bonded is cracked from resin, obtains crude residue, it is pure by reversed-phase HPLC Change, obtain fluffy white solid 100 (5.8mg, 6.2ol are fluffy, 59%, HPLC after purification with tfa salt calculate yield).
1H NMR(400MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 11.30 (s, 1H, uracil- ), NH 10.86 (s, 1H, Trp-NH), 8.60 (t, J=5.6Hz, 0.5H, CONHCH2), 8.41 (d, J=9.1Hz, 0.5H, ), CONH 8.37 (d, J=9.2Hz, 0.5H, CONH), 8.12 (t, J=5.4Hz, 0.5H, CONHCH2),8.04(br s,3H, NH3), 7.84-7.82 (m, 1H, Ar-H), 7.79-7.74 (m, 2H, Ar-H), 7.64 (m, 1H, Ar-H), 7.55 (d, J= 8.1Hz,0.5H,H-6),7.52-7.50(m,1.5H,1x Ar-H+0.5H x H-6),7.47-7.42(m,2H,Ar-H), 7.33-7.30(m,2H,Ar-H),7.11(m,1H,Ar-H),7.06-7.03(m,1H,Ar-H),6.98-6.95(m,1H,Ar- ), H 6.47-6.44 (m, 1H, NHCONH), 6.42-6.38 (m, 1H, NHCONH), 5.72 (d, J=3.7Hz, 0.5H, H-1 '), 5.69 (d, J=3.5Hz, 0.5H, H-1 '), 5.60-5.57 (m, 1H, H-5), 4.82-4.77 (m, 0.5H, DABA- β-CH), 4.61 (t, J=9.4Hz, 0.5H, DABA- α-CH), 4.55-4.48 (m, 1.5H, 0.5xDABA- α-CH+1x Trp- α-CH), 4.39-4.33(m,2.5H,0.5x H-4’+1x H-2’+1x Nal-α-CH),4.30-4.25(m,0.5H,H-4’),3.96- 3.78(m,2H,1.5x Gly-α-CH2+0.5x DABA-β-CH),3.58-3.55(m,0.5H,Gly-α-CH2),3.26-3.12 (m,2H,H-5’),3.15-3.06(m,2H,1x Trp-β-CH2+1x Nal-β-CH2), 2.99 (dd, J=14.7,6.9Hz, 1H,Nal-β-CH2),2.94-2.90(m,1H,Trp-β-CH2),2.78(s,1.5H,NCH3),2.76(s,1.5H,NCH3), 2.22-2.12 (m, 1H, H-3 '), and 1.70-1.62 (m, 1H, H-3 '), 1.14 (d, J=6.5Hz, 1.5H, CH3),1.04(d,J =6.9Hz, 1.5H, CH3)；13C NMR(100MHz,(CD3)2The 1:1 mixture of SO, ca. rotational isomer) δ 174.8, 172.1,170.3,170.0,166.7,166.3,163.9,163.8,158.9,158.7,158.4,158.2,157.6, 151.2,141.8,141.5,136.7,135.8,135.7,133.6,132.4,128.6,128.5,128.3,128.1, 128.0,127.9,127.8,126.4,125.9,124.4,124.3,121.4,119.1,119.0,116.7,111.9, 110.1,102.4,102.0,92.6,92.5,79.2,79.1,74.6,74.5,55.4,55.0,54.9,54.8,54.0, 53.9,53.6,51.1,44.5,44.3,38.8,38.6,36.3,36.1,29.4,28.4,27.9,15.2,14.4；LRMS[M+ H]+940.0..The data of these data and Tran et al. report are consistent.[4]
Assessment/Pharmacological Testing Results
Full cell inhibits test Mtb
Material and method
MtbH37Rv (ATCC 27294) and MtbH37Ra (ATCC 25177) bacterial strain are being supplemented with OADC (Difco experiment Room, Detroit, MI, USA), Middlebrook (Middlebrook) 7H9 liquid of 0.05% glycerol and 0.05% Tween-80 It is grown in culture medium.The culture being newly inoculated with is grown about 14 days at 37 DEG C, until Midexponential (OD6000.4-0.8), for pressing down System test.
The step of influence that these analogs grow MtbH37Rv by Taneja and Tyagi by previously being described2It uses The measurement of resazurin reduction microwell plate measuring method.7H9S culture medium (Middlebrook 7H9 contain 10%ADC, 0.05% glycerol, 0.05% Tween 80,1% tryptone) in, grow to Midexponential (OD600Mtb 0.4-0.8) is diluted to OD600 0.002；96 hole microtiter plates have 100 titration inhibitor, and serial dilution is into 7H9S.It is added into each hole diluted (100 μ L represent~2 × 10 to Mtb4CFU mL-1).By plate at 37 DEG C, in moist incubator, it is incubated for 5 days, then to each 0.02% resazurin solution (30 μ L) and 20% Tween-80 (12.5 μ L) are added in hole.After 24 hours, in BMG Labtech Fluorescent, excitation wavelength 530nm, launch wavelength 590nm are measured on Polarstar Omega instrument.We depict The change in fluorescence figure of negative control hole (no MtbH37Rv) is subtracted relative to Positive control wells (the not MtbH37Rv of inhibitor), To measure MIC50Value.
Shown in result such as Fig. 1 to 21 and the following table 1 of these measurements.
Table 1
Intracellular fragmentation test
Material
Rifampin is purchased from Sigma-Aldrich (R3501).All positive controls are dissolved in 100% dimethyl sulfoxide (DMSO)(0231-500mL；Amresco in), and with contain 10%ADC (BSAL；), MoregateBiotech 0.05% glycerol (GA010-P；) and 0.05% Tween 80 (P6224-500ml Chem-Supply；Sigma-Aldrich 7H9 fluid nutrient medium) (27131；DifcoBectonDickinson it) dilutes.
Neurological susceptibility test is carried out using the macrophage culture of Mtb infection
MtbH37Ra plants (ATCC25177) for infecting human macrophage like cell system (THP-1；ATCCTIB-202).Letter For it, be supplemented with 10%FBS (FBS-500；) and 0.05mM beta -mercaptoethanol (M7522 Scientifix-life； Sigma-Aldrich RPMI-1640 culture medium) (contains phenol red, 25mM HEPES and 2mM L-Glutamine；Gibco in), THP-1 original seed is set to maintain 1 × 105With 1 × 106Culture density between cell/mL.By THP-1 cell with 1 × 105Cell/ The density in hole is inoculated in tissue culturing plates with 96 hole (Costar3903；Healthy and free from worry (Corning)) in, phorbol Ethyl Myristate is added Ester (PMA；Sigma-Aldrich, 100nM).Make THP-1 cell at 37 DEG C, 5%CO2Lower differentiation 48 hours.
Cell suspending liquid of the MtbH37Ra of ultrasonic treatment in RPMI-1640 cell culture medium is used to infect differentiation THP-1 cell, at 37 DEG C, 5%CO2Lower infection 4 hours, infection multiplicity (MOI) are 5.Then supernatant is removed from all holes, With 200 μ L phosphate buffered saline (PBS) (PBS) (98-317-LB；Cellgro) THP-1 cell is washed three times, then with fresh RPMI-1640 cell culture medium, in 37 DEG C and 5%CO2Under, it is further cultured for 24 hours.
Analog is diluted in fresh RPMI-1640 cell culture medium, and is added in corresponding hole.At 37 DEG C, 5% CO2After culture 72 hours, the tissue culture medium (TCM) containing test compound is removed from hole.Cell is washed with 200 μ LPBS, then With the sterile water (T7253 containing 0.1%TritonX；Sigma-Aldrich it) cracks.Cell lysate is continuously dilute with 1:10 It releases, and in 1/10000 diluted Middlebrook 7H11/OADC (283010；Difco) bed board on agar.Then by agar Plate is cultivated 3-4 weeks at 37 DEG C, enumeration of bacterial colonies and measures CFU/mL cell lysate later.
The intracellular anti-mycobacteria activity of analog 25,36 and 37 is as shown in figs. 22-24.
The screening of the growth inhibition of one group of Gram-positive and gramnegative bacterium bacterial strain
Selectivity counting screening:
The biology of screening includes: bacillus subtilis 168, aurococcus, methicillin-resistant staphylococcus Portugal Grape coccus (MRSA), staphylococcus epidermis, Yi Shi Li Site bacterium, enterococcus faecium, Escherichia coli, comma bacillus, mouse typhus sramana Salmonella, pseudomonas aeruginosa, artificial tuberculosis yersinia genus produce alkali Providence, human pallid bacillus, clostridium perfringen, Bao Graceful acinetobacter calcoaceticus.
Material and method
Use Wong et al.3The method being previously reported have evaluated analog to 15 kinds of clinically relevant Gram-positives and leather The effect of gram-negative bacteria bacterial strain.Specifically, screening group forms (BSL1: withered grass bud by six gram positive bacterial strains Spore bacillus 168, staphylococcus epidermis [ATCC14990], enterococcus faecium [ATCC6569], Yi Shi Li Site bacterium [BAA-139]； BSL2: staphylococcus aureus [ATCC29213], methicillin-resistant staphylococcus aureus (MRSA) [BAA-44] and 9 kinds of leather Lan Shi negative strain (BSL1: e. coli k12 [BW25113], Acinetobacter bauamnnii [NCIMB12457], clostridium perfringen [ATCC35029], human pallid bacillus [ATCC49687] produce alkali Providence [ATCC9886]；BSL2: pseudoconcretion Yale Gloomy Salmonella [IP2666pIBI], pseudomonas aeruginosa [ATCC27835], salmonella typhimurium LT2, vibrio cholerae O 1 [biology Type ElTorA1552].
All aureus strains, Yi Shi Li Site bacterium and enterococcus faecium are in 10mL Tryptic Soy Broth (in 1L distilled water, 17g tryptone, 3g soya peptone, 2.5g dextrose, 5gNaCl and 2.5g dipotassium hydrogen phosphate；PH7.5 in) Culture.Alkali Providence is produced, human pallid bacillus, clostridium perfringen and Acinetobacter bauamnnii are in Nutrient broth Growth in (Difco, USA), and bacillus subtilis, Escherichia coli, comma bacillus, salmonella typhimurium, P. aeruginosa Bacterium and artificial tuberculosis yersinia genus Luria culture medium (in 1L distilled water, 10g tryptone, 5g yeast extract and 10gNaCl；PH7.5 growth in).By all three culture mediums at 121 DEG C high pressure sterilization 30 minutes.The culture of inoculation is existed Shaking table (200rpm；30 DEG C) in overnight incubation.
The overnight saturation cell culture of pathogenic strain is diluted to 1:1000 with fresh culture, in sterile transparent 384 30 μ L cultures are distributed in each hole of orifice plate.300nL DMSO is divided in advance using Perkin Elmer Janus MDT robot Stock solution is evaporated to be fixed in sieve plate.After inoculation, sieve plate is stacked on plate reader/oscillator (Perkin Elmer EnVision in), OD is read once every hour600Reading, continues 24 hours.By by the OD600 reading of bacterial index early period with Concentration in each hole is associated, is generated using the computer of the pure compound (finally screening concentration is 100 μM) of serial dilution Growth curve determines MIC value.
Analog is directed to the activity of one group of Gram-positive and gramnegative bacterium
Active preliminary data of the 2. analog 4-17 of table to one group of Gram-positive and gramnegative bacterium.
Active preliminary data of 3. analog of table to one group of Gram-positive and gramnegative bacterium.
MurX enzyme inhibits
Material
UDP-N- acetyl group-d-glucosamine [aminoglucose-14C(U)]([14C] UDP-GlcNAc, specific activity 300mCi/ Mmol it) is obtained from American Radio flower pattern, UDP-MurNAc- pentapeptide is purchased from BacWAN, Coventry, United Kingdom UNIVERSITY OF WARWICK (www.warwick.ac.uk/bacwan).It by UDP-MurNAc- pentapeptide is marked by danshensu described in saving according to 9.4.3 Five peptide synthesis of UDP-MurNAc-.TLC silica gel 60F254Plate is purchased from Merck (Germany).The every other chemicals used is at least Analyze it is pure, and be purchased from Sigma-Aldrich.Mtbmc26230 be New York Einstein medical college William Jacobs rich Scholar give.
Method: from Mtb mc26230 prepare film
Mtbmc26230 in 7H9 culture medium growth (be supplemented with 0.5% (v/v) oleic acid, 0.5% (w/v) albumin, 0.2% (w/v) glucose, 24 μ g/mL D-VB5 salt and 0.2% casamino acid).2mL/g cell, by washed cell It is resuspended to buffer solution A (50mM MOPS pH 7.9,5mM MgCl2, 5mM DTT, 10% glycerol (v/v)) in, and pass through use 150 probe sonication of Soniprep is broken (10 circulations, are opened for 60 seconds, are closed within 90 seconds).By full cell lysate at 4 DEG C, with 5,000 × g is centrifuged 20 minutes.By supernatant in Optima TLX ultracentrifuge (Beckman) with 100,000 × g (4 1 hour at DEG C) further centrifugation.The sediment of film enrichment washs in buffer solution A, then with 100,000 × g ultracentrifugation. The precipitating of washing is resuspended in buffer solution A, aliquot is divided into and is freezed at -80 DEG C.Use BCA Protein Assay Kit (Pierce) protein concentration of fraction of the estimation rich in film.
The preparation of red sulfonylation UDP-MurNAc pentapeptide
The synthesis and purifying for realizing UDP-MurNAc- pentapeptide are reproduced by the chemical zymetology of cell in vitro matter route of synthesis4。 The UDP-MurNAc- pentapeptide of the desalination in sterile water is mixed with isometric acetone, and under stiring, with 42 times of molar ratios Dansyl Cl reaction is overnight.Before rotary evaporation, red sulphonyl is quenched with the Tris.Cl pH 9 of 10 times of molar excess in reaction Chlorine is to remove solvent.Dry product is resuspended in 1mL sterile water, and pre-equilibrated with 1.5CV 0.1M ammonium hydrogen carbonate It is purified on 10/300 column of Superdex peptide (GE Healthcare) by size exclusion chromatography.Red sulfonylation UDP-MurNAc- Pentapeptide is first peak of elution during this, has feature absorbance at 340nm and 280nm.Required product will be contained Fraction is freeze-dried 4 times, and is resuspended in a small amount of sterile water.Quantitative and purity is confirmed by absorbance and mass spectrum respectively.
Radiochemistry MurX inhibits measurement
It measures mixture (200 μ L) and contains 50mM MOPSpH7.9,5mMMgCl2, 5mM DTT, 10% glycerol (v/v), 0.1%CHAPS, 100 μM of ATP, 25 μM of UDP-MurNAc- pentapeptides, 0.5 μM [14C]] inhibition of UDP-GlcNAc and various concentration (initial screening carries out under the single concentration of 200nM, and screens most effective compound under a series of concentration with determination for agent IC50Value).By the way that 400-500 μ g Mtbmc is added26230 memebrane protein initiation reactions, and cultivated 1 hour at 37 DEG C.6mL is added Chloroform/MeOH (2:1) terminates reaction, and then low-speed centrifugal, organic extract liquid is moved in the second pipe.Twice by extract backwash [once use water (800 μ L), then use chloroform/MeOH/ water (3:47:48)], be evaporated to dryness under nitrogen flowing, and be dissolved in chloroform/ MeOH (2:1v/v).A aliquot is subjected to liquid scintillation counting (LS6500, BeckmanCoulter)；By second part etc. Sample is divided to carry out thin-layer chromatography (the silica gel 60F in chloroform/MeOH/ water/ammonium hydroxide (88:48:10:1)254) development.Pass through phosphorus Light is imaged (TyphoonTRIO, AmershamBiosciences) and detects radioactive distribution, and uses ImageQuantTLv2005 Software (AmershamBiosciences) is quantitative.IC is calculated using GraFit software (version 5.0.13)50Value.
Analog is at 200nM to the inhibitory activity of MurX
Inhibition of the 4. analog 7-11 of table to MurX
Inhibition of the 5. analog 21-28 and 30-37 of table to MurX
5-28 referring to fig. 2.
Fluorescence-based MurX inhibits measurement: the active inhibition of analog 25,36 and 37 pair MurX
Continuous fluorescence MurX measurement, as previously described5(being modified slightly), usually 100 μ L volume, in vitro at 25 DEG C, Contain 83mM Tris pH 7.5,21mM MgCl2, 6% glycerol, 0.1%TritonX-100,15 μM of red sulfonylation UDP- MurNAc- pentapeptide, 40 μ g/mL polyisopreneyl phosphates and various concentration inhibitor composition measurement buffer in into Row.By adding 60-70 μ gMtb mc2The starting reaction of 6230 memebrane proteins, and 340 and 530nm excitation wavelength and hair are detected respectively Ejected wave is long.It is measured in duplicate, and calculates IC using GraFit software (version 5.0.13)50Value.
Referring to fig. 29 to 33.
Initial in vitro DMPK research
The stability of people and mice plasma
Method: the people collected and Switzerland's outbred mice plasma sample are thawed and be incorporated in DMSO/ acetonitrile/water (20:40: 40) the test compound solution prepared in, to provide final compound concentration as 1000ng/mL and final DMSO and acetonitrile concentration Respectively 0.2% and 0.4% (v/v).Fresh microcentrifugation is transferred to by blood plasma vortex mixed and by aliquot (50 μ L) Guan Zhong, and cultivated at 37 DEG C.In the different time points of 6 hour incubation period, duplicate plasma sample is taken out, and immediately dry It is rapidly frozen in ice.By all samples at -80 DEG C stored frozen, until LC-MS analyze.Passed through using 2 times of excessive acetonitriles Protein precipitation handles sample, is then centrifuged for.Supernatant analysis is carried out using Waters (Milford, MA) Acquity UPLC, It is coupled with Waters Micromass Quattro Premier mass spectrograph, with the positive electrospray ionisation mould of multiple-reaction monitoring Formula operation.Cone voltage is 45V, and 25,36 and 37 collision energies are respectively 30,30 and 40V, and m/z conversion is respectively 782.30 > 126.06 (25), 826.32 > 170.16 (36) and 888.39 > 126.06 (37).The sample (3 μ L) of processing is injected into On Supelco Ascentis Express RP Amide column (50x2.1mm, 2.7 μm), and (respectively contained using water/acetonitrile 0.05% formic acid) gradient elution analysis object, 4 minutes are lasted, flow velocity 0.4mL/min.By in blank people or mice plasma The calibration curve of middle preparation relatively quantifies sample concentration.
Table 6. measures 25,36,37 concentration in people and mice plasma after cultivating 6 hours at 37 DEG C
Vitro stability data of the table 7. 71,78,79 in people and mice plasma.
Data are the average value and standard deviation of the triplicate sample of each time point.
* relative to the measurement concentration calculation residue percentage of 2 minutes points
The average value of #n=2 sample；One duplicate sample is omitted as exceptional value
The stability of people and Mouse Liver Microsomes
Method: under 37 DEG C and 0.4mg/mL microsomal protein matter, by every kind of test compound (0.5 μM) in duplicate with People and Mouse Liver Microsomes (XenoTech, Kansas City, KS) culture are to assess metabolic stability.By the way that NADPH is added again Raw system causes metabolic response, and contains diazepam as interior target acetonitrile, not during culture in 60 minutes by being added It is quenched with time point.In the case where co-factor is not present, control sample (being free of NADPH) was quenched to supervise at 2,30 and 60 minutes Survey potential degradation.Centrifuge A sample, and pass through lcms analysis supernatant.It is analyzed using Waters Acquity UPLC, institute It states Waters Acquity UPLC and Waters Xevo G2 QTOF mass spectrograph is coupled, the mass spectrograph is with positive electrospray ionisation MSEMode operation, cone voltage are 30V.By sample (5 μ L) be injected into Ascentis Express Amide column (50 × 2.1mm, 2.7 μm) on, and use water/acetonitrile (containing 0.05% formic acid) gradient elution 4 minutes, flow velocity 0.4mL/min.From level-one Degradation curve determines degradation rate half-life period and external inherent clearance value.
Table 8. is in the small intracorporal stability of people and mouse liver
The stability of table 9. people and Mouse Liver Microsomes.
The MIC of other mycobacterias of table 10. 71 and 79 pair.
1.Boojamra, C.G. et al. J.Am.Chem.Soc.123,870-874 (2001).
2.Taneja,N.K.；Tyagi,J.S.J.Antimicrob.Chemother.60,288-293(2007).
3.Wong,Weng R.；Et al. Chem.Biol.19,1483-1495 (2012).
4.Lloyd, A.J. et al. J.Biol.Chem.283,6402-6417 (2008).
5.Mihalyi, A. doctoral thesis, UNIVERSITY OF WARWICK (2014).

Claims (62)

1. Formulas I compound represented
Or its salt, solvate, polymorph or prodrug；
Wherein,
For singly-bound or double bond；Wherein whenWhen for singly-bound, the spatial chemistry of the position is R- configuration；
R1Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkyl, C1- C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxy alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1- C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, fragrant amino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aralkyl Base amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
R2Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkyl, C1- C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxy alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1- C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Virtue Alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
R3For-C (O) R4Or-CH2R4；With
R4It is selected from the group: hydroxyl, or optionally replace: C1-C15Alkyl, C1-C15Alkyl amino；C1-C15Alkoxy；C1-C15Alkyl Sulfenyl；C1-C15Halogenated alkyl, C1-C15Halogenated alkoxy, C1-C15Hydroxy alkyl, C1-C15Alkyl carboxyl, C1-C15Alkyl carboxylic acyl Amine, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1-C4Alkyl) C3-C7Heterocycle, aryl, virtue Oxygroup, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkane Base) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
Collateral condition:
WhenWhen for double bond, R3It is not-COOH, and
WhenFor singly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl amino ,- CH(CH3)OH、-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2SCH3；And R2For methyl, isopropyl, different Butyl ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Substituted benzyl Benzyl, naphthalene or the-CH that base, aryl replace2When cyclohexyl, R3It is not-COOH.
2. Formula II compound represented:
Or its salt, solvate, polymorph or prodrug；
Wherein,
For singly-bound or double bond；Wherein whenWhen for singly-bound, the spatial chemistry of the position is R- configuration.
R1Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkyl, C1- C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxy alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1- C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, fragrant amino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aralkyl Base amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
R2Be selected from the group: hydrogen, halogen, sulfydryl, hydroxyl, acyl group, carboxyl, nitro, cyano optionally replace: C1-C6Alkyl, C1- C6Alkyl amino；C1-C6Alkoxy；C1-C6Alkyl sulfenyl；C1-C6Halogenated alkyl, C1-C6Halogenated alkoxy, C1-C6Hydroxy alkyl, C1-C6Alkyl carboxyl, C1-C6Alkylcarboxamide, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1- C4Alkyl) C3-C7Heterocycle, aryl, aryloxy group, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Virtue Alkyl amino, heteroaryl, (C1-C4Alkyl) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
R3For-C (O) R4Or-CH2R4；With
R4It is selected from the group: hydroxyl, or optionally replace: C1-C15Alkyl, C1-C15Alkyl amino；C1-C15Alkoxy；C1-C15Alkyl Sulfenyl；C1-C15Halogenated alkyl, C1-C15Halogenated alkoxy, C1-C15Hydroxy alkyl, C1-C15Alkyl carboxyl, C1-C15Alkyl carboxylic acyl Amine, C3-C7Naphthenic base；(C1-C4Alkyl) C3-C7Naphthenic base, C3-C7Heterocycle；(C1-C4Alkyl) C3-C7Heterocycle, aryl, virtue Oxygroup, arylamino, arylthio, C1-C4Aralkyl, C1-C4Aralkoxy, C1-C4Aryl alkyl amino, heteroaryl, (C1-C4Alkane Base) heteroaryl, amino, carbamoyl, amino-sulfonyl, urea groups and aroyl；
Collateral condition:
WhenWhen for double bond, R3It is not-COOH, and
WhenFor singly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl amino ,- CH(CH3)OH、-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2SCH3；And R2For methyl, isopropyl, different Butyl ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Substituted benzyl Benzyl, naphthalene or the-CH that base, aryl replace2When cyclohexyl, R3It is not-COOH.
3. compound as claimed in claim 2, wherein compound has structure shown in formula III:
Or its salt, solvate, polymorph or prodrug；
Wherein R1, R2And R4As defined in claim 2,
Collateral condition:
WhenWhen for double bond, R4It is not-OH, and
When beingSingly-bound and R1The benzyl ,-C replaced for hydrogen, methyl, isobutyl group, benzyl, fluorine or hydroxyl3-C4Alkyl amino ,- CH(CH3)OH、-CH2COOH、-CH2C(O)C(CH3)3、-CH(OH)CH3Or-CH2CH2SCH3；And R2For methyl, isopropyl, different Butyl ,-(CH2)4NH2、-CH2CH2SCH3、-CH2CH2S(O)CH3、C1-C2Aralkyl, fluorine-substituted benzyl ,-CF3Substituted benzyl Benzyl, naphthalene or the-CH that base, aryl replace2When cyclohexyl, R4It is not-OH.
4. compound as claimed in claim 2, wherein compound has formula IV:
Or its salt, solvate, polymorph or prodrug；
Wherein R1, R2And R4Definition is as claimed in claim 2.
5. compound according to any one of claims 1 to 4,For double bond.
6. compound according to any one of claims 1 to 4, wherein R1Be selected from the group: hydrogen optionally replaces: C1-C6Alkane Base, C1-C2The aralkyl, (C optionally replaced1-C2Alkyl) heteroaryl, C1-C6Alkyl amino；C1-C6Alkoxy, C1-C6Alkyl carboxylic Base, C1-C6Hydroxy alkyl.
7. compound as claimed in claim 6, wherein R1It is selected from the group: benzyl, the naphthalene, C that hydrogen, methyl, hydroxyl replace2-C4Alkyl Amino, cyclohexyl ,-CH2Cyclohexyl or-CH (OH) CH3。
8. the compound as described in any one of claims 1 to 7, wherein R2Selected from the following group group optionally replaced: C1-C6Alkane Base, (C1-C2)C3-C7Naphthenic base, C1-C4Aralkyl and C1-C6Alkyl sulfenyl.
9. compound as claimed in claim 8, wherein R2It is selected from the group: isopropyl ,-CH2Naphthalene ,-CH2Cyclohexyl ,- CH2CH2SCH3；-CH2CH2S(O)CH3Or
10. compound as claimed in any one of claims 1-9 wherein, wherein R3For-C (O) R4。
11. the compound as described in any one of claims 1 to 10, wherein R4Selected from the following group group optionally replaced: C1-C15 Alkoxy, C1-C4Aralkyl；C1-C15Alkyl amino；C1-C4Aralkoxy, C1-C4Aryl alkyl amino, C1-C15Alkyl amino.
12. compound as claimed in claim 11, wherein R4Be selected from the group: methoxyl group, hexyloxy, dodecyloxy, hydroxyl ,- CH2C(CH3)3,-O- benzyl ,-NH- benzyl ,-NH- benzyl, hexylamino.
13. compound as claimed in claim 12, wherein R4It is selected from the group :-CH2C(CH3)3,-O- benzyl ,-NH- benzyl or oneself Amino.
14. compound as claimed in claim 1 or 2, wherein the compound is selected from:
Or its salt, solvate, polymorph or prodrug.
15. compound as claimed in claim 14, wherein the compound is selected from:
Or its salt, solvate, polymorph or prodrug.
16. the compound as described in claim 1 and/or 2, wherein the compound is selected from:
Or its salt, solvate, polymorph or prodrug.
17. compound as claimed in claim 16, wherein the compound is selected from:
Or its salt, solvate, polymorph or prodrug.
18. a kind of selected from the compound comprising the following group:
Or its salt, solvate, polymorph or prodrug.
19. compound as claimed in claim 18, wherein compound is:
20. a kind of pharmaceutical composition, it includes the compound or its salt of any one of preceding claims, solvate, polycrystalline Type object or prodrug and pharmaceutically acceptable excipient.
21. a kind of method of disease or illness that prevention and/or treatment are adjusted by bacterium, the lactation including giving this needs is dynamic Compound or its salt, solvate, polymorph or its prodrug of any one of claims 1 to 19 of object therapeutically effective amount, Or composition described in claim 20.
22. method as claimed in claim 22, wherein bacterium is gram-positive bacterium.
23. the method as described in claim 21 or 22, wherein the bacterium is selected from: mycobacterium tuberculosis (Mycobacteriu M tuberculosis), mycobacterium avium (Mycobacterium avium), Mycobacterium leprae (Mycobacterium Lepra e), mycobacterium abscessus (Mycobacterium abscessus), Mycobacterium bovis (Mycobacterium Bovis), bacillus subtilis (Bacillus subtilis), staphylococcus epidermis (Staphylococcus Epidermis), enterococcus faecium (Enterococc us faecium), Yi Shi listeria spp (Listeria ivanovii), Staphylococcus aureus (Staphylococcus aureus), methicillin-resistant staphylococcus aureus (methicillin- resistant Staphylococcus aureus)。
24. a kind of prevention and/or treatment are by mycobacterium tuberculosis (Mtb) disease adjusted or the method for illness, including have given The compound or its salt, solvate, polycrystalline of this effective amount of any one of claims 1 to 19 of mammalian therapeutic needed Composition described in type object or prodrug or claim 20.
25. disease or illness that a kind of prevention and/or treatment are adjusted by the bacterium with phosphoric acid-MurNAc- pentapeptide translocase Method, including giving compound described in a effective amount of any one of claims 1 to 19 of mammalian therapeutic of this needs Or composition described in its salt, solvate, polymorph or prodrug or claim 20.
26. method as claimed in claim 25, wherein phosphoric acid-MurNAc- pentapeptide translocase is MurX.
27. the method as described in claim 21 or 22, wherein the disease or illness be selected from it is following any one: leprosy, Opportunistic infections in tuberculosis, tuberculosis, bronchiectasis and HIV/AIDS and cystic fibrosis.
28. a kind of prevent and/or treat method lungy, a effective amount of power of mammalian therapeutic including giving this needs Benefit require any one of 1-19 compound or its salt, solvate, polymorph or prodrug or claim 20 described in group Close object.
29. method as claimed in claim 28, wherein the tuberculosis is to Rimactazid, ethambutol and/or pyrrole The treatment of carboxamide dihydrochloride is resistant.
30. compound or its salt, solvate, polymorph or prodrug as described in any one of claim 1-19, or power Benefit require 20 described in composition in preparation for preventing and/or treating the use in the drug for adjusting disease or illness by bacterium On the way.
31. purposes as claimed in claim 30, wherein bacterium is gram-positive bacterium.
32. the purposes as described in claim 30 or 31, wherein the bacterium is selected from: mycobacterium tuberculosis, mycobacterium avium, fiber crops Wind mycobacteria, mycobacterium abscessus, Mycobacterium bovis, bacillus subtilis, staphylococcus epidermis, enterococcus faecium, Lee Yi Shi This special Salmonella, staphylococcus aureus, methicillin-resistant staphylococcus aureus.
33. compound or its salt, solvate, polymorph or prodrug as described in any one of claim 1-19, or power Benefit require 20 described in composition in preparation disease or illness are adjusted by mycobacterium tuberculosis (Mtb) for preventing and/or treating Drug in purposes.
34. compound or its salt, solvate, polymorph or prodrug as described in any one of claim 1-19, or power Benefit requires 20 composition to be adjusted for preventing and/or treating by the bacterium with phosphoric acid-MurNAc- pentapeptide translocase in preparation Disease or illness drug in purposes.
35. purposes as claimed in claim 34, wherein phosphoric acid-MurNAc- pentapeptide translocase is MurX or MraY.
36. the purposes as described in claim 30 or 31, wherein the disease or illness be selected from it is following any one: leprosy, Opportunistic infections in pulmonary tuberculosis, tuberculosis, bronchiectasis and HIV/AIDS and cystic fibrosis.
37. compound or its salt, solvate, polymorph or prodrug as described in any one of claim 1-19, or power Benefit require 20 described in composition preparation for preventing and/or treating the purposes in tuberculosis.
38. compound or its salt, solvate, polymorph or prodrug or root as described in any one of claim 1-19 According to the composition of claim 20, it is used to prevent and/or treat the disease or illness adjusted by bacterium.
39. compound or composition as claimed in claim 38, wherein the bacterium is gram-positive bacterium.
40. the purposes as described in claim 38 or 39, wherein the bacterium is selected from: mycobacterium tuberculosis, mycobacterium avium, fiber crops Wind mycobacteria, mycobacterium abscessus, Mycobacterium bovis, bacillus subtilis, staphylococcus epidermis, enterococcus faecium, Lee Yi Shi This special Salmonella, staphylococcus aureus, methicillin-resistant staphylococcus grape.
41. compound or its salt, solvate, polymorph or prodrug as described in any one of claim 1-19, or power Benefit require 20 described in composition, be used to prevent and/or treat by mycobacterium tuberculosis (Mtb) adjust disease or illness.
42. compound or its salt, solvate, polymorph or prodrug as described in any one of claim 1-19, or power Benefit require 20 described in composition, for prevent and/or treat by with phosphoric acid-MurNAc- pentapeptide translocase bacterium adjust Disease or illness.
43. compound as claimed in claim 42, wherein phosphoric acid-MurNAc- pentapeptide translocase is MurX or MraY
44. the compound or composition as described in claim 38 or 39, wherein the disease or illness are selected from following any one Kind: opportunistic infections and cystic fibrosis in leprosy, pulmonary tuberculosis, tuberculosis, bronchiectasis and HIV/AIDS.
45. compound or its salt, solvate, polymorph or prodrug as described in any one of claim 1-19, or power Benefit requires 20 composition, is used to prevent and/or treat tuberculosis.
46. the compound as used in claim 45, wherein tuberculosis is to Rimactazid, ethambutol and/or pyrazine The treatment of amide is resistant.
47. the method as described in any one of claim 24,33 and 41, purposes or compound, wherein Mtb is the H37Rv of Mtb Bacterial strain.
48. method as claimed in claim 21, wherein bacterium is Acinetobacter bauamnnii.
49. method as claimed in claim 21, wherein bacterium is Escherichia coli.
50. method as claimed in claim 21, wherein bacterium is pseudomonas aeruginosa.
51. method as claimed in claim 21, wherein bacterium is staphylococcus aureus.
52. method as claimed in claim 21, wherein bacterium is methicillin-resistant staphylococcus aureus.
53. method as claimed in claim 21, wherein bacterium is comma bacillus.
54. method as claimed in claim 21, wherein bacterium is clostridium perfringen.
55. method as claimed in claim 21, wherein bacterium is human pallid bacillus.
56. method as claimed in claim 21, wherein bacterium is to produce alkali Providence.
57. method as claimed in claim 21, wherein bacterium is bacillus subtilis.
58. method as claimed in claim 21, wherein bacterium is dung enterobacteria.
59. method as claimed in claim 21, wherein bacterium is Yi Shi listeria spp.
60. method as claimed in claim 21, wherein bacterium is staphylococcus epidermis.
61. method as claimed in claim 21, wherein bacterium is salmonella typhimurium.
62. method as claimed in claim 21, wherein bacterium is artificial tuberculosis yersinia genus.
CN201780086783.1A 2016-12-16 2017-12-15 The new compound of anti-mycobacteria Pending CN110300759A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000044335A2 (en) * 1999-01-28 2000-08-03 Microcide Pharmaceuticals, Inc. Uridyl peptide antibiotic (upa) derivatives, their synthesis and use
CN105755076A (en) * 2016-04-11 2016-07-13 中国医学科学院医药生物技术研究所 Method for synthesizing and obtaining SANSANMYCIN structural analogue by utilizing mutation

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000044335A2 (en) * 1999-01-28 2000-08-03 Microcide Pharmaceuticals, Inc. Uridyl peptide antibiotic (upa) derivatives, their synthesis and use
CN105755076A (en) * 2016-04-11 2016-07-13 中国医学科学院医药生物技术研究所 Method for synthesizing and obtaining SANSANMYCIN structural analogue by utilizing mutation

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
CONSTANTINE G. BOOJAMRA 等: "Synthetic Dihydropacidamycin Antibiotics: A Modified Spectrum of Activity for the Pacidamycin Class", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》 *
YUANYUAN SHI 等: "Improving the N-terminal diversity of sansanmycin through mutasynthesis", 《MICROBIAL CELL FACTORIES》 *

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