Amphotericin B semisynthetic derivatives and their preparation methods and uses

Abstract

The present invention provides semisynthetic derivatives of amphotericin B, their preparation methods, and uses. Specifically, the present invention discloses compounds having a structure as shown in Formula V, as well as water-soluble salts and complexes, pharmaceutical compositions, and plant and body therapeutic products containing the derivatives, and their use as antifungal antibiotics. The present invention also discloses methods for preparing the aforementioned compounds. #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical synthesis, and more particularly, relates to a semi-synthetic derivative of amphotericin B, a synthesis method thereof, and use thereof in treating fungal infectious diseases. Background Art

[0002] Although invasive fungal infections account for a relatively low proportion of fungal infections, they have received widespread attention due to their high mortality and morbidity rates. Currently, invasive fungal infections have become a major cause of death from major diseases such as AIDS and malignant tumors, rising to the third largest infectious disease, seriously threatening human life and causing enormous damage to human health and the economy.

[0003] Amphotericin B (AmB) was the first drug used to treat invasive fungal infections and has been used clinically since the 1950s. The advantages of amphotericin B include a broad antimicrobial spectrum, strong antimicrobial activity, bactericidal effects at high concentrations, and a low incidence of acquired resistance observed in clinical practice. It remains the last resort for treating severe invasive Aspergillus infections. However, the use of AmB is limited by its poor water solubility and severe side effects, such as hemolytic toxicity, nephrotoxicity, and infusion site reactions. Due to the toxic and side effects of amphotericin B (particularly hemolytic toxicity and nephrotoxicity), its therapeutic window is very narrow, and clinical dosages often fail to reach its optimal therapeutic concentration. For this reason, amphotericin B is currently reserved for the treatment of life-threatening invasive fungal infections.

[0004] Currently, the mortality rate for severe invasive Aspergillus infections without amphotericin B is as high as 50%, resulting in 1.5 million deaths annually. This demonstrates the critical need for low-toxicity amphotericin B for human health. Despite the tireless efforts of scientists worldwide, no highly effective and low-toxic amphotericin B derivatives are currently available. Research and development of low-toxic, high-efficiency amphotericin B drugs include developing liposomal formulations of amphotericin B, which reduce the polymer content, improve its metabolism and distribution in the body, and reduce its toxicity. Currently, three liposomal formulations of amphotericin B are on the market, but these liposomes are complex to manufacture, expensive, and, most importantly, have limited toxicity reduction. A second solution is to structurally modify amphotericin B, conducting structure-activity relationship and structure-toxicity relationship studies to control, improve, or even eliminate its toxicity at the source, thereby developing the next generation of highly active, low-toxic polyene antifungal drugs. Due to the complex structure of amphotericin B, current structural modifications are typically performed at the carboxyl terminus of the macrocycle and the amino terminus of the trehalose.

[0005] In this field, there have been many studies on new derivatives with carboxyl-terminal modifications, mainly carboxyl esterification or amidation, while trehalose amino groups are mainly modified by amidation or alkylation. Compared with amphotericin B, such derivatives have little structural change, and some are still amphiphilic. Therefore, although the overall toxicity is reduced, the activity of most is reduced several times, and the safety window has not been significantly improved. Structural modification of amphotericin B has always been a difficulty and hot spot in the research of polyene macrocyclic antibiotics. Currently, there are no new chemical entities derived from the structural modification of amphotericin B for the treatment of fungal infections. The main reason is that these new derivatives are difficult to achieve a balance between antibacterial activity, toxicity, and metabolism, and have not reached the ideal level. The therapeutic window is not substantially improved compared with amphotericin.

[0006] In summary, there is an urgent need in the art to provide new amphotericin B derivatives with novel structures, stronger antibacterial activity, and significantly reduced hemolytic activity. Summary of the Invention

[0007] To address these challenges, the present invention provides a class of diamidated amphotericin B derivatives with novel structures, enhanced antibacterial activity, and significantly reduced hemolytic activity. These compounds possess novel structures and a therapeutic index significantly superior to that of amphotericin B, potentially representing a new class of highly effective, low-toxic antifungal drugs.

[0008] The first aspect of the present invention provides a compound described in the following formula V, or a pharmaceutically acceptable salt thereof:

[0009]

[0010] in,

[0011] Each R1 is independently selected from the group consisting of: -NR4-(L) p -Ra; wherein Ra is selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C1-C6 amino, -NH2, substituted or unsubstituted -NH-C1-C 10 Alkyl, substituted or unsubstituted -NH-C(O)C1-C 10 alkyl, substituted or unsubstituted 6-10 membered aryl, substituted or unsubstituted 5-10 membered heteroaryl, or substituted or unsubstituted benzo 5-6 membered heteroaryl, and -COR3;

[0012] L is selected from the group consisting of: (CH2) n 、(CH2OCH2) m , NH, NHC (O); wherein, when each CH2 and NH appears alone or as part of another group, it may be optionally substituted;

[0013] R2 are each independently selected from the group consisting of: H, -C(O)(L) p -Rc; wherein, Rc are each independently selected from the group consisting of: (CH2) n NHR4, NHR4, or a substituted or unsubstituted 3-8 membered heterocyclic group (preferably a nitrogen-containing heterocyclic group); and when R2 is H, R1 is -NR4-(L) p -Ra, and at least one L in the R1 group is NH or NHC(O);

[0014] R3 are each independently selected from the group consisting of NH2, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted phenyl, substituted or unsubstituted five-membered or six-membered heteroaromatic group, or substituted or unsubstituted benzo five-membered or six-membered heteroaromatic group;

[0015] R4 are each independently selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl;

[0016] or R4 and (L) p -Ra, and the connected N atoms together constitute a substituted or unsubstituted 5-12 membered heterocyclic group, or a substituted or unsubstituted 5-12 membered heteroaryl group; wherein the 5-12 membered heterocyclic group or 5-12 membered heteroaryl group may have an Rb substituent, and the Rb substituent is selected from the following group: hydroxyl, unsubstituted or hydroxyl substituted C1-C 10 Alkyl, substituted or unsubstituted 5-12 membered heterocyclic group;

[0017] p is selected from the group consisting of 0, 1, 2, or 3;

[0018] n is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, or 8;

[0019] m is selected from the group consisting of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10;

[0020] Unless otherwise specified, the term "substituted" refers to one or more hydrogen atoms on the group being replaced by a substituent selected from the group consisting of halogen, oxo, carboxyl, cyano, hydroxyl, 5-6 membered heterocycle, C1-C6 alkyl, C1-C6 alkoxy, C3-C8 cycloalkyl, amino, N,N-dimethylamino, mercapto, C1-C6 sulfone, -(CH2OCH2) n OH, (CH2OCH2) n OR4, (CH2OCH2) n NH2, (CH2OCH2) n NHR4, (CH2OCH2) n N(R4)2.

[0021] In another preferred embodiment, the R1 are each independently selected from the following group: -NR4-(L) p -Ra; wherein Ra is selected from the group consisting of: -NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted 6-10 membered aryl, substituted or unsubstituted 5-10 membered heteroaryl, -COR3;

[0022] L is selected from the group consisting of: (CH2) n 、(CH2OCH2) m , NH, NHC (O); wherein, when each CH2 and NH appears alone or as part of another group, it may be optionally substituted;

[0023] p is 0 or 1;

[0024] R4 are each independently selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl;

[0025] or R4 and (L) p -Ra, and the connected N atoms together constitute a substituted or unsubstituted 5-12 membered heterocyclic group, or a substituted or unsubstituted 5-12 membered heteroaryl group; wherein the 5-12 membered heterocyclic group or 5-12 membered heteroaryl group may have an Rb substituent, and the Rb substituent is selected from the following group: hydroxyl, unsubstituted or hydroxy-substituted C1-C10 alkyl, substituted or unsubstituted 5-12 membered heterocyclic group.

[0026] In another preferred embodiment, the R2 are each independently selected from the following group: H, -C(O)(L) p -Rc; wherein, Rc are each independently selected from the following group: NHR4, or a substituted or unsubstituted 3-8 membered heterocyclic group (preferably a nitrogen-containing heterocyclic group);

[0027] L is selected from the group consisting of: (CH2) n 、(CH2OCH2) m , NH, NHC (O); wherein, when each CH2 and NH appears alone or as part of another group, it may be optionally substituted;

[0028] p is 0 or 1.

[0029] In another preferred embodiment, the n is selected from the following group: 0, 1, 2, 3, 4 or 5.

[0030] In another preferred embodiment, the m is selected from the following group: 0, 1, 2, 3, 4 or 5.

[0031] In another preferred embodiment, the R1 is selected from the following group: -NR4(CH2) n R5, -NHCH2CH2NH2,

[0032]

[0033] wherein R5 is selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl, and substituted or unsubstituted 5-12 membered heterocyclic group;

[0034] R6 is selected from the group consisting of H, OH, a substituted or unsubstituted C1-C6 alkyl group, and a substituted or unsubstituted 5-12 membered heterocyclic group.

[0035] In another preferred embodiment, the R2 is selected from the following group: -CO(CH2) n NHR4, -CO(CH2OCH2) m (CH2) n NHR4,

[0036] In another preferred embodiment, the substituted alkyl group is selected from the following group: C1-C6 alkyl, N,N-dimethylamino, mercapto, and sulfone-substituted C1-C6 alkyl.

[0037] In another preferred embodiment, the R2 is selected from the following group:

[0038]

[0039] In another preferred embodiment, the R1 is selected from the following group:

[0040]

[0041] In another preferred embodiment, the R2 is selected from the following group:

[0042]

[0043] In another preferred embodiment, or a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from the following table:

[0044]

[0045]

[0046]

[0047]

[0048] The second aspect of the present invention provides a method for preparing the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, the method comprising the steps of:

[0049]

[0050] In an inert solvent, the protecting group is removed with compound M-2 to obtain a compound of formula V;

[0051] Preferably, the method further comprises steps (1) and (2):

[0052]

[0053] (1) In an inert solvent, use AmB and compound Reaction to obtain a compound of formula M-1;

[0054]

[0055] (2) reacting M-1 with compound R1H in an inert solvent to obtain a compound of formula M-2;

[0056] Wherein, the PG is an amino protecting group; the R2' is selected from the following group: chemical bond, -C(O)(L) p -Rc'; wherein Rc' is a group formed when Rc loses a hydrogen atom; and when R2' is a chemical bond, R1 is -NR4-(L) p -Ra, and at least one L in the R1 group is NH or NHC(O);

[0057] The definitions of the remaining groups are the same as those described in the first aspect of the present invention.

[0058] In another preferred embodiment, the PG and the amino group N to which it is connected constitute a carbamate, an amide, an N-alkylamine or an N-arylamine. Preferably, the protecting group is selected from Fmoc, Boc, Cbz, Alloc, Teoc, methoxycarbonyl or ethoxycarbonyl, Tfa, Pht, Tos, Ns, pivaloyl, benzoyl, Bn, PMB, Trt, Dmb; more preferably, the protecting group is selected from the group consisting of Fmoc, Boc, Tos, and Cbz.

[0059] The third aspect of the present invention provides a method for preparing the compound according to the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, the method comprising the steps of:

[0060]

[0061] In an inert solvent, the protecting group is removed with compound M-4 to obtain a compound of formula V;

[0062] Preferably, the method further comprises steps (a) and (b):

[0063]

[0064] (a) In an inert solvent, use AmB and compound Reaction to obtain a compound of formula M-3;

[0065]

[0066] (b) reacting a compound of formula M-3 with a compound PG-R1'H in an inert solvent to obtain a compound of formula M-4;

[0067] Wherein, the PG is an amino protecting group;

[0068] The R1' is a group formed by R1 losing a H atom;

[0069] The R2' is selected from the following group: chemical bond, -C(O)(L) p -Rc'; wherein Rc' is a group formed when Rc loses a hydrogen atom; and when R2' is a chemical bond, R1 is -NR4-(L) p -Ra, and at least one L in the R1 group is NH or NHC(O);

[0070] The definitions of the remaining groups are the same as those described in the first aspect of the present invention.

[0071] The fourth aspect of the present invention provides a pharmaceutical composition comprising the following components:

[0072] 1) a therapeutically effective amount of one or more compounds of the first aspect of the present invention, or optical isomers thereof, or pharmaceutically acceptable salts, hydrates, or solvates thereof; and

[0073] 2) Pharmaceutically acceptable carriers or excipients.

[0074] The fifth aspect of the present invention provides a use of the compound described in the first aspect of the present invention, or its optical isomers, pharmaceutically acceptable salts, hydrates, and solvates, characterized in that it is used to prepare a pharmaceutical composition for preventing or treating infectious diseases caused by fungal infections.

[0075] In another preferred embodiment, the fungus is selected from the group consisting of Candida, Cryptococcus neoformans, Aspergillus (preferably Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Aspergillus niger), fungi, dermatophytes, or a combination thereof.

[0076] The sixth aspect of the present invention provides an intermediate for preparing the compound of formula V according to the first aspect of the present invention, wherein the intermediate is selected from the following group:

[0077]

[0078]

[0079] Wherein, the PG is an amino protecting group, and the amino group N connected thereto forms a carbamate, amide, N-alkylamine, or N-arylamine. Preferably, the protecting group is selected from Fmoc, Boc, Cbz, Alloc, Teoc, methoxycarbonyl or ethoxycarbonyl, Tfa, Pht, Tos, Ns, pivaloyl, benzoyl, Bn, PMB, Trt, Dmb; preferably, it is selected from the group consisting of Fmoc, Boc, Tos, and Cbz; more preferably, it is Fmoc.

[0080] The R1' is a group formed by R1 losing a H atom; R2' is selected from the following group: chemical bond, -C(O)(L) p -Rc'; wherein Rc' is a group formed when Rc loses a hydrogen atom; and when R2' is a chemical bond, R1 is -NR4-(L) p -Ra, and at least one L in the R1 group is NH or NHC(O); the definitions of the remaining groups are as described in the first aspect of the present invention.

[0081] After the intermediate compound is deprotected, the compound of formula V as described in the first aspect of the present invention is obtained.

[0082] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be listed here one by one. DETAILED DESCRIPTION

[0083] After extensive research, the inventors synthesized a series of compounds. Through antimicrobial activity screening and pharmacokinetic screening, they discovered for the first time that the compound represented by the following general formula (I) possesses strong antimicrobial activity and excellent pharmacokinetic properties, making it particularly suitable as an anti-infective treatment. Based on this, the inventors completed the present invention.

[0084] the term

[0085] In the present invention, the halogen is F, Cl, Br or I.

[0086] In the present invention, unless otherwise specified, the terms used have the general meanings commonly known to those skilled in the art.

[0087] In the present invention, the term "C1-C6 alkyl" refers to a straight or branched alkyl group having 1 to 6 carbon atoms, including but not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl, etc.; preferably ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl and tert-butyl.

[0088] In the present invention, the term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, propoxy, isopropoxy, butoxy and the like.

[0089] In the present invention, the term "C2-C6 alkenyl" refers to a straight chain or branched alkenyl group having 2 to 6 carbon atoms and containing one double bond, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl.

[0090] In the present invention, the term "C2-C6 alkynyl" refers to a straight chain or branched alkynyl group having 2 to 6 carbon atoms and containing one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl and hexynyl.

[0091] In the present invention, the term "C3-C10 cycloalkyl" refers to a cyclic alkyl group having 3 to 10 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl. The terms "C3-C8 cycloalkyl," "C3-C7 cycloalkyl," and "C3-C6 cycloalkyl" have similar meanings.

[0092] In the present invention, the term "C3-C10 cycloalkenyl" refers to a cyclic alkenyl group having 3 to 10 carbon atoms in the ring, including but not limited to cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl and cyclodecyl. The term "C3-C7 cycloalkenyl" has a similar meaning.

[0093] In the present invention, the terms "aromatic ring" and "aryl group" have the same meaning. Preferably, "aryl group" is "C6-C12 aryl group" or "C6-C10 aryl group." The term "C6-C12 aryl group" refers to an aromatic ring group having 6 to 12 carbon atoms and no heteroatoms in the ring, such as phenyl and naphthyl. The term "C6-C10 aryl group" has a similar meaning.

[0094] In the present invention, the terms "aromatic heterocycle" or "heteroaryl" have the same meaning and refer to a heteroaromatic group containing one to multiple heteroatoms. The heteroatoms referred to herein include oxygen, sulfur, and nitrogen. Examples include furyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, and the like. The heteroaryl ring may be fused to an aryl, heterocyclyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heteroaryl ring. The heteroaryl group may be optionally substituted or unsubstituted.

[0095] In the present invention, the term "3-12 membered heterocyclic group" refers to a saturated or unsaturated 3-12 membered ring group containing 1 to 3 heteroatoms selected from oxygen, sulfur and nitrogen, such as dioxolanyl. The term "3-7 membered heterocyclic group" has a similar meaning.

[0096] In the present invention, the term "substituted" refers to the replacement of one or more hydrogen atoms on a specific group with a specific substituent. The specific substituent is the substituent described above, or the substituent appearing in the embodiments. Unless otherwise specified, a substituted group may have a substituent selected from a specific group at any substitutable site of the group, and the substituent may be the same or different at each position. A cyclic substituent, such as a heterocycloalkyl, may be connected to another ring, such as a cycloalkyl, to form a spirobicyclic system, for example, the two rings having a common carbon atom. It will be understood by those skilled in the art that the combinations of substituents contemplated by the present invention are those that are stable or chemically feasible. The substituents include, but are not limited to, C1-8 alkyl, C2-8 alkenyl, C2-8 alkynyl, C3-8 cycloalkyl, 3- to 12-membered heterocyclic groups, aryl, heteroaryl, halogen, hydroxyl, carboxyl (-COOH), C1-8 aldehyde, C2-10 acyl, C2-10 ester, amino, alkoxy, C1-10 sulfonyl, etc.

[0097] In the present invention, the term "protecting group" refers to a group that protects a substituted or unsubstituted amino group at a non-reactive site in the reaction, so that the amino group N to which it is connected forms a carbamate, amide, N-alkylamine or N-arylamine, for example, Fmoc, Boc, Cbz, Alloc, Teoc, methoxycarbonyl or ethoxycarbonyl, Tfa, Pht, Tos, Ns, pivaloyl, benzoyl, Bn, PMB, Trt, Dmb, etc.

[0098] In the present invention, pharmaceutically acceptable salts refer to salts of the compounds of the present invention and inorganic or organic acids, wherein the inorganic acid is selected from hydrochloric acid, sulfuric acid, and phosphoric acid, and the organic acid is selected from chiral isomers or racemates of glutamic acid, L-glutamic acid, DL-aspartic acid, proline, alanine, citric acid, L-malic acid, tartaric acid, mandelic acid, lactic acid, succinic acid, deoxycholic acid, benzoic acid, or acetic acid, oxalic acid, maleic acid, and fumaric acid.

[0099] Pharmaceutical composition and its preparation

[0100] Another aspect of the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of one or more compounds selected from the group consisting of the compounds of the above-mentioned general formula V, pharmaceutically acceptable salts, enantiomers, diastereomers, or racemates thereof, and optionally, one or more pharmaceutically acceptable carriers, excipients, adjuvants, auxiliary materials, and / or diluents. The auxiliary materials include, for example, flavoring agents, fragrances, sweeteners, and the like.

[0101] The pharmaceutical composition provided by the present invention preferably contains an active ingredient in an amount of 1-99% by weight, preferably a compound of formula V as the active ingredient accounting for 35 wt% to 99 wt% of the total weight, with the remainder being a pharmaceutically acceptable carrier, diluent, solution or saline solution.

[0102] The compounds and pharmaceutical compositions provided by the present invention can be in various forms, such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, and can be present in suitable solid or liquid carriers or diluents and suitable sterile devices for injection or infusion.

[0103] The various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional preparation methods in the pharmaceutical field. The unit dosage of the preparation formula contains 1 mg-700 mg of the compound of formula V, preferably, the unit dosage of the preparation formula contains 5 mg-300 mg of the compound of formula V.

[0104] The compounds and pharmaceutical compositions of the present invention can be used clinically in mammals, including humans and animals, and can be administered via the oral, nasal, dermal, pulmonary, or gastrointestinal routes. Oral administration is most preferred. The most preferred daily dose is 0.1-140 mg / kg body weight, taken as a single dose, or 0.1-70 mg / kg body weight, taken in divided doses. Regardless of the route of administration, the optimal individual dose will depend on the specific treatment being used. Typically, a low dose is started and gradually increased until the most suitable dose is found.

[0105] The present invention also provides a fungal infection inhibitor, which comprises one or more compounds selected from the group consisting of the compound represented by the above-mentioned general formula V, its pharmaceutically acceptable salts, racemates, R-isomers, S-isomers or mixtures thereof, and optionally one or more pharmaceutically acceptable carriers, excipients, adjuvants, auxiliary materials and / or diluents.

[0106] The compounds and compositions of the present invention can be used to inhibit fungi and thus can be used as therapeutic drugs for diseases or conditions associated with fungal infections, such as Candida albicans, Cryptococcus neoformans, Aspergillus flavus, Aspergillus fumigatus, zygotes, dermatophytes, and the like.

[0107] Therefore, another aspect of the present invention provides the use of the compound represented by the above general formula, its pharmaceutically acceptable salt, racemate, R-isomer, S-isomer or mixture thereof in the preparation of a medicament for treating diseases associated with fungal infection, such as diseases associated with Candida albicans, Candida krusei, Candida parapsilosis, Candida tropicalis, Cryptococcus neoformans, Aspergillus fumigatus and the like.

[0108] Another aspect of the present invention provides a method for treating diseases associated with fungal infections, comprising administering to a patient in need of such treatment one or more compounds selected from the group consisting of the compounds represented by the above-mentioned general formula V, their pharmaceutically acceptable salts, racemates, R-isomers, S-isomers or mixtures thereof.

[0109] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the invention. The experimental methods in the following examples, for which specific conditions are not specified, are generally based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are calculated by weight.

[0110] In all embodiments, 1 H-NMR was recorded using a Varian Mercury 400 or Varian Mercury 600 NMR instrument, and chemical shifts were expressed in δ (ppm). Low-resolution mass spectra were measured using a Finnigan MAT95 mass spectrometer. Column chromatography used silica gel of 200-300 mesh. The abbreviations and their full names appearing in the examples are as follows (in order of appearance): FmocOSu, 9-fluorenylmethyl-N-succinimidyl carbonate; DMA, N,N-dimethylacetamide; HPLC, high performance liquid chromatography; DMSO, dimethyl sulfoxide; PyAOP, (3H-1,2,3-triazolo[4,5-B]pyridine-3-oxy)tris-1-pyrrolidinyl hexafluorophosphate; TLC, thin layer chromatography; HONSu, N-hydroxysuccinimide; DCC, dicyclohexylcarbodiimide; PyBOP, 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate; DIPEA, N,N-diisopropylethylamine; DMF, N,N-dimethylformamide.

[0111] Example 1 Synthesis of Compound V-1

[0112]

[0113] Step 1:

[0114] Amphotericin B (2.26 g, 2.446 mmol, 1 eq), FmocOSu (1.5 g, 4.4 mmol, 1.8 eq), and pyridine (0.6 mL, 7.4 mmol, 3 eq) were added sequentially to a single-necked flask containing DMA (220 mL). The flask was wrapped with aluminum foil and protected with argon. The mixture was stirred overnight at room temperature and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (1000 mL) and filtered under atmospheric pressure. The filter cake was the crude product of compound 1-2. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 6:1) to obtain compound 1-2 (1.6 g) as a light yellow solid in a yield of 56.4%. MS (ESI) m / z: 1143.3 [MH] - .HRMS(ESI):Anal.Calcd for C 62 H 82 NO 19 [MH] - :1144.5487,Found:1144.5468.

[0115] Step 2:

[0116] Compound 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), acetylhydrazine (23 mg, 0.314 mmol, 2 eq), and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) were added sequentially to a single-necked vial containing DMA (5 mL). The vial was wrapped with aluminum foil and protected with argon. The mixture was stirred overnight at room temperature and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (100 mL) and filtered to obtain the crude product of compound 1-3. This was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10:1 to 5:1) to obtain compound 1-3 (100 mg) as a light yellow solid in a yield of 53%. MS (ESI) m / z: 1200.5 [MH] - .

[0117] Step 3:

[0118] Compound 1-3 (100 mg, 0.083 mmol) and piperidine (0.1 mL) were added sequentially to a single-necked flask containing DMSO (3 mL). The outer wall of the single-necked flask was wrapped with aluminum foil and protected with argon. The reaction was stirred at room temperature for 2 hours, and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (100 mL) and filtered. The resulting solid was washed with methyl tert-butyl ether (40 mL x 2), filtered, and dried under vacuum to obtain the target product V-1 (52 mg) as a light yellow solid in a yield of 64%. 1 H NMR (600MHz, DMSO-d6) δ6.51-6.03(m,12H),5.96(dd,J=15.2,8.9Hz,1H),5.88(s,1H),5.46-5.39(m,2H),5.23(m,1H),4.82- 4.77(m,2H),4.76(d,J=3.8Hz,1H),4.70(d,J=4.9Hz,1H),4.67(d,J=4.9Hz,1H),4.49-4.42(m,2H),4.35(m,1H),4.29-4.20(m ,2H),4.09-4.00(m,2H),3.56-3.45(m,3H),3.17(m,1H),2.85(m,1H),2.33-2.25(m,2H),2.19-2.14(m,2H),2.01-1.88(m,3H) ,1.84(s,3H),1.72(m,1H),1.64-1.37(m,7H),1.37-1.20(m,6H),1.15-1.09(m,7H),1.07-1.02(m,3H),0.92(d,J=7.1Hz,1H). 13 C NMR (126MHz, DMSO-d6) δ171.63,171.06,169.11,137.33,137.22,134.37,134.17,133.94,133.67,132.9 6,132.92,132.67,132.36,132.31,131.75,130.11,128.81,97.60,96.73,77.70,74.72,74.35,74.02,7 3.96,72.96,70.24,69.56,69.26,68.32,66.64,65.54,65.21,56.79,49.20,47.15,46.64,45.27,45.06 ,42.94,42.47,40.98,39.49,35.59,29.51,20.90,18.98,18.58,17.41,12.58.MS(ESI)m / z:980.5[M+H]+ .HRMS(ESI):Anal.Calcd forC 49 H 78 N3O 17 [M+H] + :980.5331,Found:980.5304.

[0119] Example 2 Synthesis of Compound V-2

[0120]

[0121] Step 1:

[0122] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), ethoxyacylhydrazine (28 mg, 0.314 mmol, 2 eq) and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) as raw materials, compound 2-1 (90 mg) was synthesized as a light yellow solid.

[0123] Step 2:

[0124] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 2-1 (100 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-2 (41 mg) as a light yellow solid. 1H NMR(600MHz,DMSO-d6)δ6.48-6.03(m,12H),5.97(m,1H),5.84(s,1H),5.45-5.37(m,2H),5.21(m,1H),4.85-4.72(m,4H),4.69-4.61( m,2H),4.50-4.41(m,2H),4.35(m,1H),4.30-4.18(m,2H),4.08-3.96(m,2H),3.58(s,2H),3.54-3.43(m,2H),3.23(m,1H),3.12-3.02 (m,3H),2.90(d,J=32.0Hz,1H),2.84(m,1H),2.74(d,J=31.8Hz,1H),2.41(m,1H),2.32-2.24(m,2H),2.19-2.12(m,2H),1.94(s,1H), 1.92-1.85(m,2H),1.70(m,1H),1.61-1.34(m,6H),1.35-1.18(m,4H),1.14-1.05(m,7H),1.02(d,J=5.8Hz,3H),0.90(d,J=6.7Hz,3H). 13 C NMR(126MHz,DMSO d6)δ172.11,171.05,157.28,137.53,137.22,134.42,134.19,134.02,133.68,132.93,132.86,132.65,132 .59,132.37,132.20,131.73,128.64,97.59,96.60,77.68,74.50,74.35,74.05(2C,overlap),72.92,70.38 ,69.59,69.30,68.30,66.66,65.56,65.15,56.85,55.31,52.32,46.69,45.33,45.24,42.93,42.49,40.92, 40.08,36.45,35.58,29.51,18.96,18.59,17.42,12.56.MS(ESI)m / z:996.5[M+H]+.HRMS(ESI):Anal.Calcd for C 49 H 78 N3O 18 [M+H]+:996.5280,Found:996.5266.

[0125] Example 3 Synthesis of Compound V-3

[0126]

[0127] Step 1:

[0128] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), aminoacylhydrazine (24 mg, 0.314 mmol, 2 eq) and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) as raw materials, compound 3-1 (85 mg) was synthesized as a light yellow solid.

[0129] Step 2:

[0130] Referring to the synthesis method of step 3 in Example 1, using DMSO as solvent, 3-1 (85 mg) and piperidine (0.1 mL) as raw materials, the target product V-3 (30 mg) was synthesized as a light yellow solid. 1 H NMR(500MHz,DMSO-d6)δ6.51-6.05(m,12H),5.95(m,1H),5.87(s,1H),5.45(m,1H),5.37(s,1H),5.23(m,1H),4.95(m,1H),4. 83-4.77(m,2H),4.75(m,1H),4.72-4.62(m,2H),4.49-4.40(m,2H),4.35(m,1H),4.30-4.21(m,2H),4.12-3.97(m,2H),3.60- 3.43(m,3H),3.19(m,1H),3.16-3.04(m,3H),2.87(m,1H),2.75(m,1H),2.44(m,1H),2.29(m,1H),2.22-2.13(m,2H),2.00-1. 87(m,2H),1.73(m,1H),1.64-1.39(m,7H),1.37-1.22(m,5H),1.18-1.08(m,7H),1.05(d,J=5.5Hz,3H),0.92(d,J=6.4Hz,3H). 13C NMR(126MHz,DMSO-d6)δ172.34,171.04,159.48,137.45,137.24,134.40,134.19,134.02,133.69,132.92,13 2.86,132.61,132.36,132.21,131.69,130.11,128.95,97.61,97.19,77.65,74.71,74.23,74.07,73.92,73. 09,70.26,69.62,69.36,68.18,66.67,65.85,65.38,56.70,55.18,46.68,45.27,44.89,42.92,42.52,40.92 ,40.07,36.94,35.56,29.47,18.96,18.54,17.45,12.55.MS(ESI)m / z:981.2[M+H]+.HRMS(ESI):Anal.Calcd for C 48 H 77 N4O 17 [M+H]+:981.5284,Found:981.5300.

[0131] Example 4 Synthesis of Compound V-4

[0132]

[0133] Step 1:

[0134] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), phenylacylhydrazine (43 mg, 0.314 mmol, 2 eq) and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) as raw materials, compound 4-1 (102 mg) was synthesized as a light yellow solid.

[0135] Step 2:

[0136] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 4-1 (102 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-4 (36 mg) as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ7.94(d,J=7.1Hz,2H),7.56(m,1H),7.51-7.42(m,2H),6.53-6.07(m,12H),6.02(m,1H),5.88(s,1H),5.49-5.37(m,2H),5.24(m,1H),4.90-4.70(m,4H),4.69-4.61(m,2H),4.52-4.41(s,2H),4.36-4.23(m,2H),4.14-4.03(m,2H),3.60-3.46(m,4H),3.17-3.04(m,3H),2.91(m,1H),2.79(m,1H),2.65(m,1H),2.36-2.25(m,2H),2.23-2.15(m,2H),2.04(m,1H),1.94(m,1H),1.73(m,1H),1.68-1.47(m,6H),1.48-1.23(m,4H),1.18(d,J=5.0Hz,3H),1.16-1.08(m,4H),1.05(d,J=5.2Hz,3H),0.93(d,J=6.1Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ171.81,171.05,166.11,137.58,137.22,134.43,134.19,134.05,133.68,132.93,132.87,132.80,132.66,132.60,132.37,132.20,132.09,131.73,128.80(2C,overlap),128.75,128.12(2C,overlap),97.61,96.67,77.68,74.63,74.33,74.06,73.53,73.01,70.07,69.61,69.31,68.29,66.67,65.63,65.29,56.63,55.60,46.72,45.32,45.18,42.92,42.49,40.92,40.03,36.73,35.59,29.52,18.96,18.60,17.43,12.56.MS(ESI)m / z:1042.3[M+H]+.HRMS(ESI):Anal.Calcd for C 54 H 80 N3O 17 [M+H]+:1042.5488,Found:1042.5466.

[0137] Example 5 Synthesis of Compound V-5

[0138]

[0139] Step 1:

[0140] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), pyridine-3-acylhydrazine (43 mg, 0.314 mmol, 2 eq) and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) as raw materials, compound 5-1 (100 mg) was synthesized as a light yellow solid.

[0141] Step 2:

[0142] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 5-1 (100 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-5 (53 mg) as a light yellow solid. 1 H NMR (500MHz, DMSO-d6) δ9.07(s,1H),8.73(d,J=3.2Hz,1H),8.27(d,J=7.2Hz,1H),7.50(m,1H),6.53-6.06(m,12H),6.02(m,1H),5.89 (s,1H),5.50-5.38(m,2H),5.23(m,1H),4.88-4.72(m,5H),4.68(m,1H),4.61(m,1H),4.51-4.40(m,2H),4.36-4.24(m,3H),4.15-4.0 1(m,2H),3.64-3.46(m,3H),3.18-3.01(m,3H),2.88(m,1H),2.35-2.23(m,2H),2.18(d,J=4.7Hz,2H),2.05(m,1H),1.94(m,1H),1.73 (m,1H),1.67-1.48(m,5H),1.47-1.22(m,7H),1.17(d,J=5.5Hz,3H),1.15-1.08(m,4H),1.05(d,J=5.3Hz,3H),0.92(d,J=6.3Hz,3H). 13C NMR (126MHz, DMSO-d6) δ171.74,171.05,164.74,152.78,149.09,137.50,137.22,135.80,134.42,134.19,134.02,133. 68,132.94,132.87,132.66,132.62,132.37,132.24,131.73,128.76,128.49,123.97,97.62,96.71,77.69,74.61,74.3 4,74.06,73.78,73.05,70.20,69.60,69.31,68.30,66.66,65.57,65.28,56.77,55.57,46.70,45.31,45.20,42.92,42. 49,40.90,40.02,36.70,35.57,29.52,18.96,18.60,17.42,12.55.MS(ESI)m / z:1043.3[M+H]+.HRMS(ESI):Anal.Calcd for C 53 H 79 N4O 17 [M+H]+:1043.5440,Found:1043.5432.

[0143] Example 6 Synthesis of Compound V-6

[0144]

[0145] Step 1:

[0146] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), pyrazinyl acylhydrazine (43 mg, 0.314 mmol, 2 eq) and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) as raw materials, compound 6-1 (86 mg) was synthesized as a light yellow solid.

[0147] Step 2:

[0148] Referring to the synthesis method of step 3 in Example 1, using DMSO as solvent and 6-1 (86 mg) and piperidine (0.1 mL) as raw materials, the target product (31 mg) was synthesized as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ9.19(m,1H),8.89(m,1H),8.75(m,1H),6.52-6.06(m,12H),6.03(m,1H),5.89(s,1H),5.49-5.39(m,2H) ,5.23(m,1H),4.85-4.73(m,3H),4.69(m,1H),4.60(m,1H),4.51-4.39(m,2H),4.35-4.24(m,2H),4.14-4.03(m,2H),3.60-3.47 (m,3H),3.16-3.06(m,3H),2.85(m,1H),2.34-2.24(m,2H),2.18(d,J=5.3Hz,2H),2.04(m,1H),1.95(m,1H),1.79(m,1H),1.73( m,1H),1.66-1.48(m,5H),1.47-1.21(m,7H),1.20-1.15(m,3H),1.14-1.08(m,4H),1.05(d,J=6.0Hz,3H),0.93(d,J=6.8Hz,3H). 13 CNMR(126MHz,DMSO-d6)δ173.44,171.04,162.50,148.16,145.08,144.27,143.98,137.49,137.22,134.43,134 .20,134.05,133.69,132.92,132.86,132.65,132.61,132.37,132.24,131.72,128.80,97.61,96.84,77.67,74. 72,74.64,74.32,74.07,73.11,70.27,69.60,69.31,68.28,66.67,65.57,65.32,56.88,55.68,46.72,45.31,4 5.04,42.94,42.50,40.91,40.04,36.75,35.58,29.55,18.97,18.60,17.43,12.55.MS(ESI)m / z:1044.3[M+H]+.

[0149] Example 7 Synthesis of Compound V-7

[0150]

[0151] Step 1:

[0152] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), furan-2-acylhydrazine (40 mg, 0.314 mmol, 2 eq) and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) as raw materials, compound 7-1 (112 mg) was synthesized as a light yellow solid.

[0153] Step 2:

[0154] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 7-1 (112 mg) and piperidine (0.1 mL) were used as raw materials to synthesize the target product V-7 (45 mg) as a light yellow solid. 1 H NMR(500MHz,DMSO-d6)δ7.86(m,1H),7.21(m,1H),6.64(m,1H),6.54-6.06(m,12H),6.00(m,1H),5.88(s,1H),5.51-5.35(m,2H),5.24( m,1H),4.84-4.74(m,3H),4.73-4.68(m,2H),4.66(m,1H),4.60(m,1H),4.46(m,1H),4.41(m,1H),4.33-4.24(m,2H),4.17(m,1H),4.13- 4.02(m,2H),3.59-3.45(m,3H),3.17-3.05(m,2H),2.84(m,1H),2.36-2.24(m,2H),2.18(d,J=4.6Hz,2H),2.01(m,1H),1.94(m,1H),1. 73(m,1H),1.67-1.46(m,5H),1.45-1.22(m,6H),1.16(d,J=5.3Hz,3H),1.14-1.08(m,4H),1.05(d,J=5.4Hz,3H),0.93(d,J=6.4Hz,3H). 13C NMR (126MHz, DMSO-d6) δ171.86,171.05,157.78,146.78,146.03,137.59,137.22,134.42,134.20,134.04,133.68,132 .93,132.86,132.66,132.59,132.37,132.19,131.72,128.72,115.19,112.27,97.61,96.89,77.68,74.72,74.29,74. 12,74.07,72.96,70.42,69.61,69.33,68.25,66.67,65.69,65.28,56.63,55.56,46.72,45.31,45.14,42.92,42.50,4 0.91,40.03,36.78,35.58,29.50,18.97,18.63,17.43,12.56.MS(ESI)m / z:1032.3[M+H]+.HRMS(ESI):Anal.Calcdfor C 52 H 78 N3O 18 [M+H]+:1032.5280,Found:1032.5256.

[0155] Example 8 Synthesis of Compound V-8

[0156]

[0157] Step 1:

[0158] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 1-2 (180 mg, 0.157 mmol, 1 eq), PyAOP (130 mg, 0.25 mmol, 1.6 eq), hydrazine (32 mg, 0.314 mmol, 2 eq) and N-methylmorpholine (47.6 mg, 0.471 mmol, 3 eq) as raw materials, compound 8-1 (91 mg) was synthesized as a light yellow solid.

[0159] Step 2:

[0160] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 8-1 (91 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-8 (28 mg) as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ6.52-6.02(m,12H),5.96(m,1H),5.84(s,1H),5.50-5.34(m,2H),5.21(m,1H),4.85 -4.73(m,3H),4.69(m,1H),4.46(m,1H),4.42-4.33(m,2H),4.32-4.20(m,2H),4.13-4.00(m,2H),3.63-3.4 5(m,3H),3.17-3.05(m,3H),2.85(m,1H),2.39-2.25(m,3H),2.22-2.13(m,2H),1.98-1.87(m,2H),1.73(m, 1H),1.68-1.37(m,8H),1.36-1.20(m,6H),1.18-1.08(m,7H),1.05(d,J=4.8Hz,3H),0.92(d,J=6.0Hz,3H). 13 C NMR (126MHz, DMSO-d6) δ173.22,171.02,160.72,158.13,137.55,137.24,134.39,134.20,134.00,133.72,132. 93,132.86,132.66,132.62,132.38,132.22,131.65,129.14,97.56,97.39,77.58,74.71,74.33,74.09,73.43, 71.81,69.70,69.41,69.09,68.27,66.71,65.35,64.99,56.79,55.63,46.81,45.39,45.11,42.91,42.51,40.9 1,40.03,36.72,35.57,29.53,18.98,18.57,17.46,12.54.MS(ESI)m / z:1009.2[M+H]+.HRMS(ESI):Anal.Calcd for C 49 H 77 N4O 18 [M+H]+:1009.5233,Found:1009.5213.

[0161] Example 9 Synthesis of Compound V-9

[0162]

[0163] Step 1:

[0164] Referring to the synthesis method of Step 1 in Example 1, amphotericin B (2.26 g, 2.446 mmol, 1 eq), 9-1 (1.8 g, 4.4 mmol, 1.8 eq; Tetrahedron. 2000, 56, 8119-8131), and pyridine (0.6 mL, 7.4 mmol, 3 eq) were added sequentially to a single-necked flask containing DMA (220 mL). The flask was wrapped with aluminum foil and protected with argon. The mixture was stirred overnight at room temperature and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (1000 mL) and filtered under atmospheric pressure. The filter cake was the crude product of compound 9-2. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 6:1) to obtain compound 9-2 (1.2 g) as a light yellow solid. MS (ESI) m / z: 1215.4 [MH] - HRMS(ESI):Anal.Calcd for C 65 H 87 N2O 20 [MH] - :1215.5858,Found:1215.5859.

[0165] Step 2:

[0166] Referring to the synthesis method of Step 2 in Example 1, compound 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), methylacylhydrazine (24 mg, 0.328 mmol, 2 eq), and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) were added sequentially to a single-necked flask containing DMA (5 mL). The outer wall of the single-necked flask was wrapped with aluminum foil and protected with argon. The mixture was stirred at room temperature overnight and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (100 mL) and filtered to obtain the crude product of compound 9-3. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10:1 to 5:1) to obtain compound 9-3 (101 mg) as a light yellow solid.

[0167] Step 3:

[0168] Referring to the synthesis method of Step 3 in Example 1, compound 9-3 (101 mg) and piperidine (0.1 mL) were added sequentially to a single-necked vial containing DMSO (3 mL). The outer wall of the single-necked vial was wrapped with aluminum foil and protected with argon. The reaction was stirred at room temperature for 2 hours, and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (100 mL) and filtered. The resulting solid was washed with methyl tert-butyl ether (40 mL x 2), filtered, and dried in vacuo to obtain the target product V-9 (31 mg) as a light yellow solid. 1 H NMR(600MHz,DMSO-d6)δ6.51-6.05(m,12H),5.96(dd,J=15.1,8.9Hz,1H),5.89(s,1H),5.43(m,2H),5.23(m,1H),4.81-4 .77(m,1H),4.76(m,1H),4.69(m,1H),4.66-4.60(m,2H),4.57(m,1H),4.47(m,1H),4.35(m,1H),4.30-4.22(m,2H),4.10 -3.99(m,2H),3.70(m,1H),3.57-3.44(m,3H),3.04(d,J=7.3Hz,2H),2.32-2.23(m,4H),2.19-2.13(m,2H),1.98-1.86(m ,4H),1.72(m,1H),1.64-1.36(m,7H),1.35-1.19(m,6H),1.16-1.06(m,7H),1.04(d,J=5.9Hz,3H),0.92(d,J=6.9Hz,3H). 13C NMR(126MHz,DMSO-d6)δ171.65,171.23,171.06,169.53,137.23,137.07,134.36,134.17,133.89,133.67,132.97,1 32.95,132.71,132.40,132.37,131.75,130.11,128.96,97.61,96.40,77.69,75.08,74.33,74.02,72.97,72.03,69. 68,69.55,69.25,68.31,66.64,65.59,65.21,55.33,54.92,49.21,46.64,45.35,45.10,43.00,42.47,40.91,39.50 ,36.82,36.69,35.59,29.50,20.86,18.99,18.53,17.41,12.58.MS(ESI)m / z:1051.6[M+H]+.HRMS(ESI):Anal.Calcd for C 52 H 83 N4O 18 [M+H]+:1051.5702,Found:1051.5714.

[0169] Example 10 Synthesis of Compound V-10

[0170]

[0171] Step 1:

[0172] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 3-amino-1,2-propanediol (30 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 10-1 (96 mg) was synthesized as a light yellow solid.

[0173] Step 2:

[0174] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 10-1 (96 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-10 (22 mg) as a light yellow solid. 1H NMR(600MHz,DMSO-d6)δ6.47-6.01(m,12H),5.93(m,1H),5.81(s,1H),5.46-5.38(m,1H),5.34(s,1H),5.19(m,1H),4.83-4.60( m,7H),4.43(m,1H),4.38(m,1H),4.33(m,1H),4.25-4.16(m,2H),4.08-3.97(m,2H),3.70(m,1H),3.56(m,1H),3.54-3.47(m,2H ),3.44(m,1H),3.15(m,1H),3.13-3.00(m,6H),2.31-2.21(m,3H),2.19-2.12(m,2H),2.01-1.90(m,2H),1.85(m,1H),1.70(m,1 H),1.60-1.34(m,7H),1.32-1.18(m,5H),1.13(d,J=4.2Hz,3H),1.11-1.05(m,4H),1.02(d,J=5.4Hz,3H),0.90(d,J=6.3Hz,3H). 13 CNMR(126MHz,DMSO-d6)δ172.86,171.53,171.04,137.34,137.26,134.39,134.18,133.99,133.69,132.93,132.88,13 2.66,132.36,132.30,131.68,130.11,129.13,97.56,97.47,77.64,75.70,75.56,74.18,74.07,73.56,71.03,70.95, 69.76,69.64,69.36,68.15,66.68,65.92,65.18,64.30,64.13,57.26,54.86,54.50,46.77,45.25,45.04,42.88,42.5 3,40.92,37.33,36.61,35.56,29.46,18.97,18.54,17.46,12.55.MS(ESI)m / z:1068.3[M+H]+.HRMS(ESI):Anal.Calcd for C 53 H 86 N3O 19 [M+H] + :1068.5856,Found:1068.5853.

[0175] Example 11 Synthesis of Compound V-11

[0176]

[0177] Step 1:

[0178] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 4-hydroxypiperidine (33 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 11-1 (97 mg) was synthesized as a light yellow solid.

[0179] Step 3:

[0180] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 11-1 (96 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-11 (24 mg) as a light yellow solid. 1 H NMR(500MHz,DMSO-d6)δ6.50-6.05(m,12H),5.97(m,1H),5.86(s,1H),5.48-5.37(m,2H),5.23(m,1H),4.83-4.69(m,6H),4.65( m,1H),4.46(m,1H),4.41-4.22(m,4H),4.12-3.97(m,3H),3.88(m,1H),3.76-3.67(m,2H),3.62(m,1H),3.57-3.45(m,2H),3.21 -2.99(m,5H),2.95(m,1H),2.83-2.75(m,2H),2.35-2.25(m,3H),2.18(m,1H),1.97(m,1H),1.89(m,1H),1.81(m,1H),1.73(m,1 H),1.64-1.38(m,8H),1.37-1.21(m,6H),1.21-1.15(m,4H),1.12(d,J=5.6Hz,3H),1.05(d,J=5.9Hz,3H),0.92(d,J=6.8Hz,3H). 13CNMR(126MHz,DMSO-d6)δ171.28,171.18,171.04,137.25,136.88,134.34,134.15,133.85,133.66,132.99,132.95,132.7 5,132.70,132.47,132.35,131.68,129.12,97.68,97.01,77.66,75.06,74.86,74.24,74.04,73.75,70.96,69.69,69.61, 69.32,68.21,66.66,66.24,54.89,54.43,51.67,51.59,46.69,45.29,45.03,43.52,42.91,42.45,40.92,40.06,37.87,3 6.60,35.57,35.40,35.00,34.64,29.49,18.98,18.54,17.43,12.55.MS(ESI)m / z:1078.3[M+H]+.HRMS(ESI):Anal.Calcd forC 55 H 88 N3O 18 [M+H]+:1078.6063,Found:1078.6075.

[0181] Example 12 Synthesis of Compound V-12

[0182]

[0183] Step 1:

[0184] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 1-(2-hydroxyethyl)piperazine (43 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 12-1 (97 mg) was synthesized as a light yellow solid.

[0185] Step 2:

[0186] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 12-1 (97 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-12 (26 mg) as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ6.51-6.05(m,12H),5.96(dd,J=14.9,9.1Hz,1H),5.87(s,1H),5.49-5.37(m,2H),5.23(m,1H),4.8 2-4.63(m,6H),4.47(m,1H),4.39-4.29(m,2H),4.26(m,1H),4.11-3.97(m,2H),3.69(m,1H),3.67-3.58(m,3H),3.56-3.43 (m,6H),3.20-3.02(m,6H),2.43-2.38(m,2H),2.33-2.25(m,3H),2.18(d,J=5.3Hz,2H),1.93-1.81(m,2H),1.73(m,1H),1. 64-1.38(m,7H),1.37-1.23(m,5H),1.17(d,J=5.5Hz,3H),1.15-1.09(m,4H),1.05(d,J=5.7Hz,3H),0.92(d,J=6.6Hz,3H). 13 C NMR (126MHz, DMSO-d6) δ171.39,171.20,171.04,137.26,136.82,134.34,134.15,133.85,133.66,132.99,132.95,132.75, 132.70,132.47,132.34,131.70,129.22,97.68,96.93,77.65,75.01,74.25,74.05,73.82,70.88,69.71,69.61,69.32,68. 22,66.67,66.56,66.22,60.61,58.96,54.91,54.46,54.27,53.82,51.47,46.79,45.99,45.28,45.03,42.86,42.50,41.77 ,40.92,40.05,36.76,36.61,35.56,29.49,18.98,18.56,17.43,12.55.MS(ESI)m / z:1107.3[M+H]+.HRMS(ESI):Anal.Calcd for C 56 H 91 N4O 18 [M+H]+:1107.6328,Found:1107.6308.

[0187] Example 13 Synthesis of Compound V-13

[0188]

[0189] Step 1:

[0190] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 1-(3-hydroxypropyl)piperazine (47 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 13-1 (102 mg) was synthesized as a light yellow solid.

[0191] Step 2:

[0192] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 13-1 (102 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-13 (32 mg) as a light yellow solid. 1 H NMR(600MHz,DMSO-d6)δ6.51-6.00(m,12H),5.93(m,1H),5.86(s,1H),5.50-5.30(m,2H),5.20 (m,1H),4.89-4.57(m,6H),4.45(m,1H),4.38-4.16(m,4H),4.10-3.91(m,2H),3.74-3.55(m,4 H),3.54-3.39(m,5H),3.17-2.92(m,6H),2.40-2.18(m,6H),2.17-2.04(m,3H),1.94-1.77(m, 2H),1.70(m,1H),1.64-1.44(m,7H),1.42-1.20(m,8H),1.20-0.94(m,13H),0.93-0.77(m,3H). 13C NMR(151MHz,DMSO-d6)δ171.35,171.11,171.02,137.20,136.77,134.33,134.10,133.76,133.57,132.98,132.94,132.74,1 32.68,132.51,132.34,131.65,129.16,97.63,96.89,77.62,74.97,74.20,73.98,73.78,70.75,69.72,69.58,69.20,68.13, 66.62,66.53,66.10,59.66,55.37,54.80,54.36,53.90,53.51,51.48,46.67,45.82,45.23,44.93,42.87,42.43,41.79,40.8 4,36.70,39.97,36.54,35.51,29.99,29.47,18.96,18.53,17.38,12.52.MS(ESI)m / z:1121.3[M+H]+.HRMS(ESI):Anal.Calcd for C 57 H 93 N4O 18 [M+H]+:1121.6485,Found:1121.6487.

[0193] Example 14 Synthesis of Compound V-14

[0194]

[0195] Step 1:

[0196] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 1-(2,2-dimethyl-2-hydroxyethyl)piperazine (52 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 14-1 (95 mg) was synthesized as a light yellow solid.

[0197] Step 2:

[0198] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 14-1 (102 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-14 (28 mg) as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ6.51-6.03(m,12H),5.96(m,1H),5.86(s,1H),5.45(m,1H),5.38(s,1H),5.22(m,1H),4.84-4 .67(m,6H),4.63(m,1H),4.45(m,1H),4.38-4.29(m,3H),4.25(m,1H),4.16-3.97(m,3H),3.70(m,1H),3.66-3.56(m, 3H),3.55-3.41(m,4H),3.21-2.97(m,7H),2.28(br,4H),2.24-2.11(m,4H),1.94-1.79(m,2H),1.73(m,1H),1.67-1. 47(m,5H),1.48-1.22(m,8H),1.18(d,J=4.7Hz,3H),1.15-1.07(m,4H),1.05(d,J=4.8Hz,3H),0.92(d,J=5.8Hz,3H). 13 C NMR(126MHz,DMSO-d6)170.85,170.67,170.54,136.76,136.33,133.83,133.65,133.36,133.16,132.49,132.45,132.24,132.2 1,131.96,131.84,131.19,128.72,97.18,96.48,77.16,74.52,73.73,73.55,73.32,72.04,70.39,70.22,69.24,69.11,68.83,6 8.32,67.70,66.17,65.71,55.29,54.94,54.40,53.94,50.92,48.70,46.26,45.70,44.78,44.48,42.00,41.67,40.42,39.51,3 6.23,36.11,35.06,28.98,28.06,27.96,26.81,18.47,18.06,16.93,12.05.MS(ESI)m / z:1135.3[M+H]+.HRMS(ESI):Anal.Calcd forC 58 H 95 N4O 18 [M+H]+:1135.6636,Found:1135.6636.

[0199] Example 15 Synthesis of Compound V-15

[0200]

[0201] Step 1:

[0202] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 1-(1,1-dimethyl-2-hydroxyethyl)piperazine (52 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 15-1 (100 mg) was synthesized as a light yellow solid.

[0203] Step 2:

[0204] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 15-1 (100 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-15 (31 mg) as a light yellow solid. 1 H NMR(500MHz,DMSO-d6)δ6.49-6.01(m,12H),5.94(m,1H),5.85(s,1H),5.47-5.35(m,2H),5.21(m,1H),4.81-4. 63(m,7H),4.44(m,1H),4.37-4.19(m,4H),4.09-3.95(m,2H),3.68(m,1H),3.34-3.55(s,3H),3.54-3.41(m,4H ),3.17-3.00(m,6H),2.79(m,1H),2.26(br,4H),2.20-2.12(m,2H),1.90-1.77(m,2H),1.71(m,1H),1.63-1.42 (m,6H),1.42-1.20(m,7H),1.15(d,J=5.0Hz,3H),1.12-1.06(m,4H),1.02(d,J=6.0Hz,3H),0.95-0.87(m,6H). 13C NMR (126MHz, DMSO-d6) δ170.69,170.65,170.54,136.76,136.26,133.84,133.66,133.35,133.16,132.50,132.45,132.25,132.21,132.01, 131.85,131.21,128.73,97.18,96.37,77.15,74.47,73.77,73.55,73 .37,70.40,69.21,69.11,68.81,67.73,66.98,66.17,66.03,65.66,65 .25,56.95,55.16,54.77,54.35,53.98,51.00,50.93,48.71,46.46,4 6.24,45.90,45.64,45.31,44.78,44.49,42.43,42.17,42.00,40.42, 39.51,36.20,36.07,35.06,28.99,26.81,20.71,20.59,18.49,18.07,16.93,12.05.MS(ESI)m / z:1135.2[M+H]+.HRMS(ESI):Anal.Calcdfor C 58 H 95 N4O 18 [M+H]+:1135.6636,Found:1135.6641.

[0205] Example 16 Synthesis of Compound V-16

[0206]

[0207] Step 1:

[0208] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 4-(4-piperidinyl)morpholine (56 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 16-1 (112 mg) was synthesized as a light yellow solid.

[0209] Step 2:

[0210] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 16-1 (112 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-16 (38 mg) as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ6.51-6.05(m,12H),5.97(m,1H),5.86(s,1H),5.50-5.35(m,2H),5.23(m,1H),4.86-4.68(m,6H),4.64(m,1H),4.49(m 1H),4.45(m,1H),4.37(m,1H),4.35-4.21(m,3H),4.13(m,1H),4.10-3.95(m,2H),3.72(m,1H),3.65-3.44(m,7H),3.19-3.03(m,6H),2.96(m,1H),2.49-2.35(m,6H),2.34-2.24(m,4H),2.22-2.14(m,2H),1.94-1.81(m,2H),1.80-1.68(m,2H),1.68-1.47(m,5H),1.46-1.22(m,9H),1.21-1.14(m,3H),1.12(d,J=5.7Hz,3H),1.05(d,J=5.6Hz,3H),0.92(d,J=6.5Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ171.23,171.04,170.58,137.27,136.81,134.32,134.15,133.83,133.65,133.00,132.97,132.77,132.71,132.49,132.35,131.69,129.27,97.68,96.98,77.66,74.97,74.28,74.04,73.81,70.83,69.70,69.62,69.34,68.23,66.99(2C,overlap),66.67,66.41,66.06,61.54,54.55,54.39,51.51,49.99,49.89,46.74,45.29,45.00,42.91,42.50,41.06,40.91,39.52,36.75,36.27,35.57,29.49,29.23,29.00,28.22,18.98,18.55,17.43,12.55.MS(ESI)m / z:1147.2[M+H]+.HRMS(ESI):Anal.Calcd for C 59 H 95 N4O 18 [M+H]+:1147.6641,Found:1147.6619.

[0211] Example 17 Synthesis of Compound V-17

[0212]

[0213] Step 1:

[0214] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 9-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 2,6-dimethyl-1-(2-hydroxyethyl)piperazine (52 mg, 0.328 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as raw materials, compound 17-1 (102 mg light yellow solid) was synthesized.

[0215] Step 2:

[0216] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 17-1 (102 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-17 (29 mg) as a light yellow solid. 1 H NMR(500MHz,DMSO-d6)δ6.52-6.04(m,12H),5.98(m,1H),5.88(s,1H),5.49-5.36(m,2H),5.23(m,1H),4.87-4.73(m,4H),4.72-4.64(m,2H),4 .47(m,1H),4.42-4.34(m,2H),4.32-4.22(m,3H),4.07(m,1H),4.00(m ,1H),3.69(m,1H),3.61(m,1H),3.57-3.49(m,3H),3.17-3.02(m,7H),2 .84(m,1H),2.71-2.59(m,3H),2.40(m,1H),2.34-2.22(m,5H),2.21-2 .12(m,2H),1.97-1.85(m,2H),1.74(m,1H),1.66-1.46(m,6H),1.46-1. 22(m,8H),1.20-1.15(m,3H),1.14-1.08(m,6H),1.07-0.98(m,7H),0.92(d,J=6.2Hz,3H).MS(ESI)m / z:1135.2[M+H]+.HRMS(ESI):Anal.Calcd forC 58 H 95 N4O 18 [M+H]+:1135.6641,Found:1135.6624.

[0217] Example 18 Synthesis of Compound V-18

[0218]

[0219] Step 1:

[0220] Referring to the synthesis method of step 1 in Example 1, compound 18-2 (1.02 g) was synthesized as a pale yellow solid using DMA as the solvent and amphotericin B (2.26 g, 2.446 mmol, 1 eq), 18-1 (1.7 g, 4.4 mmol, 1.8 eq; Chemical Science. 2014, 5, 2747-2753), and pyridine (0.6 mL, 7.4 mmol, 3 eq) as the starting materials. MS (ESI) m / z: 1201.5 [MH] - HRMS(ESI):Anal.Calcd for C 64 H 85 N2O 20 [MH] - :1201.5701,Found:1201.5685.

[0221] Step 2:

[0222] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 18-2 (200 mg, 0.166 mmol, 1 eq), PyAOP (139 mg, 0.266 mmol, 1.6 eq), 1-(2-hydroxyethyl)piperazine (42 mg, 0.332 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.498 mmol, 3 eq) as raw materials, compound 18-3 (115 mg) was synthesized as a light yellow solid.

[0223] Step 3:

[0224] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 18-3 (115 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-18 (36 mg) as a light yellow solid. 1H NMR(600MHz,DMSO-d6)δ6.50-6.01(m,12H),5.95(m,1H),5.84(s,1H),5.46-5.39(m,1H),5.37(s,1H),5.20(m,1H) ,4.80-4.65(m,5H),4.61(m,1H),4.42(m,1H),4.36-4.18(m,3H),4.09-3.95(m,2H),3.71-3.56(m,3H),3.55-3.38 (m,5H),3.17-3.01(m,6H),2.40-2.22(m,7H),2.15(s,2H),1.99(m,1H),1.91-1.78(m,2H),1.71(m,1H),1.61-1.4 3(m,6H),1.41-1.19(m,8H),1.15(d,J=4.7Hz,3H),1.11-1.06(m,4H),1.02(d,J=5.0Hz,3H),0.90(d,J=4.9Hz,3H). 13 C NMR (126MHz, DMSO-d6) δ172.92,171.38,171.05,137.26,136.80,134.32,134.14,133.83,133.66,132.98,132.77,132.7 0,132.49,132.35,131.96,131.71,129.23,97.67,96.94,77.65,74.99,74.24,74.03,73.79,71.00,69.63,69.57,69.31, 68.21,66.65,66.56,66.21,60.61,58.88,54.64,54.28,53.81,51.45,46.71,45.89,45.31,44.97,42.92,42.49,41.83,4 0.90,39.99,36.66,35.52,29.42,25.59,18.98,18.56,17.43,12.56.MS(ESI)m / z:1093.2[M+H]+.HRMS(ESI):Anal.Calcd for C 55 H 89 N4O 18 [M+H] + :1093.6172,Found:1093.6192.

[0225] Example 19 Synthesis of Compound V-19

[0226]

[0227] Step 1:

[0228] 19-1 (1.88 mL, 13 mmol), tetrabutylammonium hydrogen sulfate (440 mg, 1.3 mmol), and sodium hydroxide solution (5 M, 4 mL) were added to a single-necked flask containing dichloromethane / water (1:1, 40 mL). Then, 19-2 (1 mL, 6.46 mmol) was added dropwise to the reaction mixture. After stirring at room temperature for 5 hours, tert-butyl bromoacetate (1.88 mL, 13 mmol) was added to the reaction mixture. Stirring was continued at room temperature overnight, and the reaction progress was monitored by TLC. After the reaction was complete, dichloromethane was added to dilute the mixture to 100 mL. The organic layer was washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and purified by column chromatography (petroleum ether / ethyl acetate = 5:1) to obtain 19-3 (1.37 g) as a colorless oil. MS (ESI) m / z: 276.2 [M+H] + .

[0229] Step 2:

[0230] Dissolve 19-3 (13.7 g, 0.0498 mol) in dichloromethane (40 mL) and stir in an ice-water bath. Add trifluoroacetic acid (57 g, 0.498 mmol) dropwise to the solution. After addition, stir at room temperature for 2 hours and monitor the reaction progress by TLC. After the reaction is complete, add toluene (30 mL x 3) to the reaction solution and remove the trifluoroacetic acid by azeotropic distillation to obtain crude 19-4 (14 g) as a colorless oil, which is used directly in the next reaction. MS (ESI) m / z: 118 [MH] - .

[0231] Step 3:

[0232] The crude product 19-4 was dissolved in water and solid sodium carbonate was added until the pH of the solution was between 8 and 9. A solution of FmocOSu (20 g, 60.7 mmol) in 1,4-dioxane (30 mL) was then added dropwise to the reaction mixture. After addition, the mixture was stirred overnight at room temperature and the reaction progress monitored by TLC. After completion of the reaction, the organic solvent was removed by concentration under reduced pressure. The remaining aqueous phase was adjusted to a pH of 8-9 with solid sodium carbonate and then extracted with ethyl acetate (20 mL x 3). The aqueous phase was retained and the pH of the solution was adjusted to 2-3 with 6M dilute hydrochloric acid. The solution was extracted again with ethyl acetate (20 mL x 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent, yielding 19-5 (13.2 g) as a white solid. MS (ESI) m / z: 340 [MH] - .

[0233] Step 4:

[0234] 19-5 (1 g, 2.93 mmol), HONSu (0.4 g, 3.52 mmol), and DCC (0.73 g, 3.52 mmol) were added sequentially to a single-necked flask containing dichloromethane (50 mL). The mixture was stirred at room temperature overnight, and the reaction progress was monitored by TLC. After the reaction was complete, the filtrate was filtered and concentrated under reduced pressure to remove the solvent. The residue was diluted to 100 mL with ethyl acetate, resulting in the appearance of a white solid. The filtrate was filtered again and washed sequentially with saturated sodium bicarbonate solution, water, and saturated brine, dried over anhydrous sodium sulfate, filtered, and purified by column chromatography to obtain 19-6 (0.66 g) as a white solid. MS (ESI) m / z: 439.2 [M+H] + .

[0235] Step 5:

[0236] Referring to the synthesis method of step 1 in Example 1, using DMA as solvent, amphotericin B (2.26 g, 2.446 mmol, 1 eq), 19-6 (1.9 g, 4.4 mmol, 1.8 eq), and pyridine (0.6 mL, 7.4 mmol, 3 eq) as raw materials, compound 19-7 (1.3 g) was synthesized as a light yellow solid. MS (ESI) m / z: 1245.4 [MH] - .HRMS(ESI):Anal.Calcd forC 66 H 89 N2O 21 [MH] - :1245.5963,Found:1245.5959.

[0237] Step 6:

[0238] Referring to the synthesis method of step 2 in Example 1, using DMSO as solvent and 19-7 (200 mg, 0.166 mmol, 1 eq), PyAOP (139 mg, 0.266 mmol, 1.6 eq), 1-(2-hydroxyethyl)piperazine (42 mg, 0.332 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.498 mmol, 3 eq) as raw materials, compound 19-8 (103 mg) was synthesized as a light yellow solid.

[0239] Step 7:

[0240] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 19-8 (103 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-19 (33 mg) as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ6.50-6.04(m,12H),5.96(m,1H),5.88(s,1H),5.48-5.36(m,2H),5.22(m,1H),4.85-4.61(m,6 H),4.47(m,1H),4.40-4.28(m,3H),4.25(m,1H),4.04(m,2H),3.91(s,2H),3.72(m,1H),3.69-3.58(m,3H),3.21-3.05( m,5H),2.89-2.77(m,2H),2.43-2.36(m,2H),2.35-2.25(m,2H),2.21-2.13(m,2H),1.93-1.79(m,2H),1.73(m,1H),1. 64-1.46(m,6H),1.45-1.22(m,7H),1.20-1.14(m,3H),1.13-1.09(m,4H),1.04(d,J=4.4Hz,3H),0.92(d,J=5.7Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ171.39,171.03,169.37,137.26,136.81,134.34,134.16,133.86,133.66,132.97,132.94,132.74,1 32.69,132.47,132.31,131.69,129.22,97.68,96.86,77.65,75.00,74.23,74.05,73.85,71.87,71.69,70.32,70.17,69.60, 69.52,69.32,68.20,66.66,66.56,66.23,60.61,58.95,54.79,54.26,53.84,51.48,46.67,45.94,45.27,44.91,42.86,42.5 1,41.83,40.92,40.60,36.69,35.63,29.47,18.98,18.59,17.43,12.55.MS(ESI)m / z:1137.4[M+H]+.HRMS(ESI):Anal.Calcd for C 57 H 93 N4O 19 [M+H]+:1137.6434.Found:1137.6460.

[0241] Example 20 Synthesis of Compound V-20

[0242]

[0243] Step 1:

[0244] Referring to the synthesis method of step 1 in Example 1, compound 20-2 (1.3 g) was synthesized using DMA as solvent and amphotericin B (2.26 g, 2.446 mmol, 1 eq), 20-1 (1.8 g, 4.4 mmol, 1.8 eq; WO2014068443), and pyridine (0.6 mL, 7.4 mmol, 3 eq) as raw materials. MS (ESI) m / z: 1215.9 [MH] - HRMS(ESI):Anal.Calcd forC 65 H 87 N2O 20 [MH] - :1215.5858,Found:1215.5839.

[0245] Step 2:

[0246] Referring to the synthesis method of step 2 in Example 1, compound 20-3 (97 mg) was synthesized as a light yellow solid using DMA as the solvent and 20-2 (200 mg, 0.164 mmol, 1 eq), PyAOP (137 mg, 0.262 mmol, 1.6 eq), 1-(2-hydroxyethyl)piperazine (43 mg, 0.328 mmol, 2 eq), and N-methylmorpholine (50 mg, 0.492 mmol, 3 eq) as the starting materials. MS (ESI) m / z: 1329.0 [M+H] + .

[0247] Step 3:

[0248] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 20-3 (103 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-20 (27 mg) as a light yellow solid. 1H NMR(600MHz,DMSO-d6)δ6.48-6.02(m,12H),5.93(dd,J=14.9,8.9Hz,1H),5.84(s,1H),5.42(dd,J=14.9,10.2Hz,1H),5.32(m,1H),5.20(m,1H),4.79-4.75m,2H),4.73(d,J=3.7Hz,1H),4.68(d,J=5.8Hz,1H),4.61(d,J=5.1Hz,1H),4.47-4.39(m,2H),4.34-4.27(m,2H),4.23(m,1H),4.08-3.95(m,3H),3.69-3.55(m,4H),3.53-3.40(m,6H),3.14-3.05(m,5H),2.84-2.74(m,2H),2.41-2.35(m,4H),2.35-2.22(m,6H),2.19-2.12(m,2H),2.02-1.92(m,2H),1.90-1.77(m,3H),1.71(m,1H),1.64-1.37(m,11H),1.34-1.18(m,9H),1.18-1.12(m,3H),1.11-1.06(m,4H),1.02(d,J=6.2Hz,3H),0.90(d,J=7.0Hz,3H). 13 C NMR(126MHz,DMSO-d6)δ171.72,171.38,171.05,137.25,136.90,134.33,134.14,133.85,133.66,132.96,132.73,132.44,132.35,131.71,130.11,129.17,128.51,97.68,96.90,77.60,74.95,74.27,74.08,73.90,70.50,69.56,69.31,68.20,66.65,66.60,66.26,60.61,58.91,55.12,54.31,53.77,51.43,49.16,46.75,45.89,45.29,45.02,42.96,42.43,41.77,40.06,38.46,37.89,35.55,29.48,27.26,25.52,19.01,18.69,17.40,12.52.MS(ESI)m / z:1107.5[M+H]+.HRMS(ESI):Anal.Calcd for C 56 H 91 N4O 18[M+H]+:1107.6328.Found:1107.6328.

[0249] Example 21 Synthesis of Compound V-21

[0250]

[0251] Step 1:

[0252] Referring to the synthesis method of step 1 in Example 19, using dichloromethane / water as solvent, 19-1 (5 mL, 0.0342 mmol) and 21-1 (3 g, 0.0171 mmol) as raw materials, 21-2 (2.7 g) was obtained as a colorless oil. MS (ESI) m / z: 290.2 [M+H] + .

[0253] Step 2:

[0254] Referring to the synthesis method of step 2 in Example 19, compound 21-3 (5 g) was obtained as a colorless oil using dichloromethane as solvent and 21-2 (5 g, 0.0173 mol) as starting material. MS (ESI) m / z: 132 [MH] - .

[0255] Step 3:

[0256] Referring to the synthesis method of step 3 in Example 19, using water / 1,4-dioxane as solvent and 21-3 (5 g) as raw material, 21-4 (7 g) was obtained as a colorless oil. MS (ESI) m / z: 354 [MH] - .

[0257] Step 4:

[0258] Referring to the synthesis method of step 4 in Example 19, compound 21-5 (3.5 g) was obtained as a colorless oil using dichloromethane as the solvent and compound 21-4 (7 g) as the starting material. MS (ESI) m / z: 452.2 [M+H]+.

[0259] Step 5:

[0260] Referring to the synthesis method of step 1 in Example 1, using DMA as solvent, amphotericin B (2.26 g, 2.446 mmol, 1 eq), 21-5 (2 g, 4.4 mmol, 1.8 eq), and pyridine (0.6 mL, 7.4 mmol, 3 eq) as raw materials, compound 21-6 (1.12 g) was synthesized as a light yellow solid. MS (ESI) m / z: 1259.4 [MH] - .HRMS(ESI):Anal.Calcd for C 67 H91 N2O 21 [MH] - :1259.6120,Found:1259.6134.

[0261] Step 6:

[0262] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 21-6 (200 mg, 0.158 mmol, 1 eq), PyAOP (132 mg, 0.254 mmol, 1.6 eq), 1-(2-hydroxyethyl)piperazine (41 mg, 0.316 mmol, 2 eq) and N-methylmorpholine (48 mg, 0.474 mmol, 3 eq) as raw materials, compound 21-7 (104 mg) was synthesized as a light yellow solid.

[0263] Step 7:

[0264] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 21-7 (104 mg) and piperidine (0.1 mL) as raw materials, and the target product V-21 (31 mg) was synthesized as a light yellow solid. 1 H NMR(500MHz,DMSO-d6)δ6.51-6.03(m,12H),5.96(m,1H),5.87(s,1H),5.50-5.34(m,2H),5.23(m,1H),4.85-4.59(m,6H) ,4.50-4.41(m,2H),4.39-4.29(m,3H),4.26(m,1H),4.11-3.97(m,2H),3.93-3.86(m,2H),3.72(m,1H),3.67-3.60(m,2H) ,3.59-3.44(m,6H),3.19-3.04(m,4H),2.78-2.66(m,2H),2.42-2.24(m,8H),2.22-2.10(m,2H),1.94-1.79(m,2H),1.73( m,1H),1.65-1.38(m,8H),1.36-1.22(m,5H),1.21-1.15(m,3H),1.14-1.08(m,4H),1.07-1.00(m,3H),0.97-0.88(m,3H). 13C NMR (126MHz, DMSO-d6) δ171.38,171.04,169.43,137.26,136.81,134.33,134.15,133.84,133.66,132.97,132.95,132.73,132 .70,132.47,132.35,131.69,129.21,97.67,96.89,77.70,75.02,74.23,74.05,73.84,70.31,70.24,69.59,69.56,69.32,68. 20,66.66,66.57,66.22,60.61,58.96,54.64,54.26,53.88,51.48,50.61,46.65,45.95,45.21,45.05,43.02,42.57,41.93,40 .92,40.07,36.83,35.92,35.63,29.40,27.23,18.97,18.59,17.43,12.55.MS(ESI)m / z:1151.6[M+H]+.HRMS(ESI):Anal.Calcd forC 58 H 95 N4O 19 [M+H]+:1151.6591.Found:1151.6601.

[0265] Example 22 Synthesis of Compound V-22

[0266]

[0267] Step 1:

[0268] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 18-2 (200 mg, 0.166 mmol, 1 eq), PyAOP (139 mg, 0.266 mmol, 1.6 eq), 2,6-dimethyl-1-(2-hydroxyethyl)piperazine (53 mg, 0.332 mmol, 2 eq) and N-methylmorpholine (50 mg, 0.498 mmol, 3 eq) as raw materials, compound 22-1 (103 mg) was synthesized as a light yellow solid.

[0269] Step 2:

[0270] Referring to the synthesis method of step 3 in Example 1, using DMSO as solvent, 22-1 (104 mg) and piperidine (0.1 mL) as raw materials, the target product V-22 (28 mg) was synthesized as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ6.51-6.03(m,12H),5.95(m,1H),5.86(s,1H),5.48-5.35(m,2H),5.21(m,1H),4.84-4.72(m,4H),4.71-4.60( m,2H),4.46(m,1H),4.40-4.32(m,2H),4.31-4.20(m,3H),4.10-3.93(m,3H),3.72-3.56(m,3H),3.55-3.43(m,2H),3.19-3.01(m,5H) ,2.70-2.58(m,2H),2.33-2.21(m,2H),2.19-2.09(m,2H),1.93-1.82(m,2H),1.78-1.67(m,2H),1.65-1.44(m,5H),1.43-1.20(m,8H) ,1.19-1.13(m,3H),1.12-1.08(m,6H),1.07-0.96(m,7H),0.91(d,J=4.8Hz,3H).MS(ESI)m / z:1121.1[M+H]+.HRMS(ESI):Anal.Calcd for C 57 H 93 N4O 18 [M+H]+:1121.6485,Found:1121.6488.

[0271] Example 23 Synthesis of Compound V-23

[0272]

[0273] Step 1:

[0274] Referring to the synthesis method of step 1 in Example 1, compound 23-2 (1.32 g) was synthesized as a pale yellow solid using DMA as the solvent and amphotericin B (2.26 g, 2.446 mmol, 1 eq), 23-1 (1.9 g, 4.4 mmol, 1.8 eq; CN106589069), and pyridine (0.6 mL, 7.4 mmol, 3 eq). MS (ESI) m / z: 1241.5 [MH] - .HRMS(ESI):Anal.Calcdfor C 67 H 89 N2O 20 [MH] - :1241.6014,Found:1241.6002.

[0275] Step 2:

[0276] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 23-2 (200 mg, 0.16 mmol, 1 eq), PyAOP (134 mg, 0.257 mmol, 1.6 eq), 2,6-dimethyl-1-(2-hydroxyethyl)piperazine (51 mg, 0.32 mmol, 2 eq) and N-methylmorpholine (49 mg, 0.48 mmol, 3 eq) as raw materials, compound 23-3 (106 mg) was synthesized as a light yellow solid.

[0277] Step 3:

[0278] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 23-4 (106 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-23 (25 mg) as a light yellow solid. 1 H NMR(500MHz,DMSO-d6)δ6.52-6.04(m,12H),5.97(m,1H),5.86(s,1H),5.49-5.36(m,2H),5.23(m,1H),4.86-4.73(m,4H),4.72-4.61(m, 2H),4.46(m,1H),4.42-4.34(m,2H),4.33-4.21(m,3H),4.11-3.94(m,3H),3.68-3.44(m,6H),3.19-3.02(m,5H),2.87(m,1H),2.81-2.72 (m,2H),2.71-2.59(m,3H),2.34-2.23(m,2H),2.22-2.12(m,2H),2.04-1.84(m,3H),1.82-1.69(m,3H),1.68-1.48(m,7H),1.49-1.23(m, 8H),1.21-1.14(m,4H),1.13-1.10(m,4H),1.09-0.99(m,7H),0.92(d,J=6.2Hz,3H).MS(ESI)m / z:1161.4[M+H]+.HRMS(ESI):Anal.Calcd for C 60 H 97 N4O 18 [M+H]+:1161.6798.Found:1161.6774.

[0279] Example 24 Synthesis of Compound V-24

[0280]

[0281] Step 1:

[0282] Referring to the synthesis method of step 3 in Example 19, using water / 1,4-dioxane as solvent and 24-1 (1.5 g) as raw material, 24-2 (3.8 g) was obtained as a colorless oil. MS (ESI) m / z: 350 [MH] - .

[0283] Step 2:

[0284] Referring to the synthesis method of step 4 in Example 19, using dichloromethane as solvent and 24-2 (3.8 g) as raw material, 24-3 (3.2 g) was obtained as a colorless oil. MS (ESI) m / z: 449.1 [M+H] +

[0285] Step 3:

[0286] Referring to the synthesis method of step 1 in Example 1, using DMA as solvent, amphotericin B (2.26 g, 2.446 mmol, 1 eq), 24-3 (1.97 g, 4.4 mmol, 1.8 eq), and pyridine (0.6 mL, 7.4 mmol, 3 eq) as raw materials, compound 24-4 (1.25 g) was synthesized as a light yellow solid. MS (ESI) m / z: 1255.6 [MH] - .HRMS(ESI):Anal.Calcd forC 68 H 91 N2O 20 [MH] - :1255.6171,Found:1255.6139.

[0287] Step 4:

[0288] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 24-4 (200 mg, 0.16 mmol, 1 eq), PyAOP (134 mg, 0.257 mmol, 1.6 eq), 2,6-dimethyl-1-(2-hydroxyethyl)piperazine (51 mg, 0.32 mmol, 2 eq) and N-methylmorpholine (49 mg, 0.48 mmol, 3 eq) as raw materials, compound 24-5 (95 mg) was synthesized as a light yellow solid.

[0289] Step 5:

[0290] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 24-5 (106 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-24 (23 mg) as a light yellow solid. 1H NMR(500MHz,DMSO-d6)δ6.50-6.04(m,12H),5.96(m,1H),5.87(s,1H),5.49-5.36(m,2H),5.23(m,1H),4.83-4.71(m,4H),4.70-4.61(m,2 H),4.45(m,1H),4.42-4.34(m,2H),4.33-4.21(m,3H),4.11-3.95(m,3H),3.71-3.51(m,2H),3.56-3.45(m,2H),3.19-3.04(m,5H),2.94(m ,1H),2.70-2.59(m,3H),2.34-2.22(m,2H),2.18(d,J=5.3Hz,2H),1.94-1.85(m,2H),1.83-1.66(m,4H),1.65-1.44(m,7H),1.44-1.23(m, 10H),1.22-1.15(m,3H),1.14-1.10(m,4H),1.09-0.98(m,9H),0.92(d,J=6.8Hz,3H).MS(ESI)m / z:1175.4[M+H]+.HRMS(ESI):Anal.Calcd for C 61 H 99 N4O 18 [M+H]+:1175.6954.Found:1175.6957.

[0291] Example 25 Synthesis of Compound V-25

[0292]

[0293] Step 1:

[0294] Dissolve 25-1 (2.64 mL, 0.03076 mol) in acetonitrile (30 mL) and stir in an ice-water bath. Add 19-1 (1.5 mL, 0.01025 mol) dropwise to the above solution. After addition, stir at room temperature overnight and monitor the reaction progress by TLC. After the reaction is complete, concentrate under reduced pressure to remove the solvent. The residue is diluted to 100 mL with dichloromethane, washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent to obtain crude 25-2 (1 g) as a colorless oil. MS (ESI) m / z: 174.2 [M+H]+.

[0295] Step 2:

[0296] Crude product 25-2 (10 g, 57.8 mmol) was dissolved in dichloromethane (40 mL) and stirred in an ice-water bath. Trifluoroacetic acid (43 mL, 578 mmol) was added dropwise to the solution. After addition, the mixture was stirred at room temperature for 2 hours and the reaction progress was monitored by TLC. After the reaction was complete, toluene (30 mL x 3) was added to the reaction solution and the trifluoroacetic acid was removed by azeotropic distillation to obtain 19.4 g of crude product as a colorless oil. The crude product was dissolved in water and solid sodium carbonate was added to adjust the solution pH to 8-9. A solution of FmocOSu (20 g, 60.7 mmol) in 1,4-dioxane (30 mL) was added dropwise to the solution. After addition, the mixture was stirred at room temperature overnight and the reaction progress was monitored by TLC. After the reaction was complete, the organic solvent was removed by concentration under reduced pressure. The remaining aqueous phase was adjusted to a pH of 8-9 with solid sodium carbonate and then extracted with ethyl acetate (20 mL x 3). The aqueous phase was retained and the pH of the solution was adjusted to 2-3 with 6M dilute hydrochloric acid. The solution was extracted again with ethyl acetate (20 mL x 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to remove the solvent to obtain 25-3 (7.6 g) as a solid. MS (ESI) m / z: 340.2 [M+H]+

[0297] Step 3:

[0298] Referring to the synthesis method of step 4 in Example 19, compound 25-4 (8.1 g) was obtained as a white solid using dichloromethane as the solvent and 25-3 (7.6 g) as the starting material. MS (ESI) m / z: 437.2 [M+H]+.

[0299] Step 4:

[0300] Referring to the synthesis method of step 1 in Example 1, using DMA as solvent, amphotericin B (2.26 g, 2.446 mmol, 1 eq), 25-4 (1.97 g, 4.4 mmol, 1.8 eq), and pyridine (0.6 mL, 7.4 mmol, 3 eq) as raw materials, compound 25-5 (1.1 g) was synthesized as a light yellow solid. MS (ESI) m / z: 1243.5 [MH] - .HRMS(ESI):Anal.Calcd forC 67 H 91 N2O 20 [MH] - :1243.6171,Found:1243.6163.

[0301] Step 5:

[0302] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent and 25-5 (200 mg, 0.16 mmol, 1 eq), PyAOP (134 mg, 0.257 mmol, 1.6 eq), 2,6-dimethyl-1-(2-hydroxyethyl)piperazine (51 mg, 0.32 mmol, 2 eq) and N-methylmorpholine (49 mg, 0.48 mmol, 3 eq) as raw materials, compound 25-6 (97 mg) was synthesized as a light yellow solid.

[0303] Step 6:

[0304] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 25-6 (97 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-25 (26 mg). 1 H NMR(500MHz,DMSO-d6)δ6.50-6.06(m,12H),5.96(m,1H),5.87(s,1H),5.49-5.36(m,2H),5.23(m,1H),4.82-4.71(m,4H),4.70-4.62(m,2H ),4.45(m,1H),4.42-4.22(m,5H),4.11-3.94(m,3H),3.69(m,1H),3. 60(m,1H),3.57-3.45(m,2H),3.44-3.36(m,2H),3.18-3.04(m,6H),2. 71-2.59(m,3H),2.36(m,1H),2.33-2.21(m,2H),2.18(d,J=5.5Hz,2H),2.05-1.85(m,2H),1.74(m,1H),1.66-1.38(m,7H),1.38-1.14(m,9 H),1.14-1.10(m,3H),1.10-1.01(m,8H),0.99(d,J=6.1Hz,6H),0.93(d,J=6.8Hz,3H).MS(ESI)m / z:1163.4[M+H]+.HRMS(ESI):Anal.Calcd for C 60 H 99 N4O 18 [M+H]+:1163.6954.Found:1163.6943.

[0305] Example 26 Synthesis of Compound V-26

[0306]

[0307] Step 1:

[0308] Referring to the synthesis method of step 2 in Example 1, using DMA as solvent, 20-2 (200 mg), PyAOP (137 mg), 1-(3-hydroxypropyl)piperazine (44 mg) and N-methylmorpholine (49 mg) as raw materials, compound 26-1 (102 mg) was synthesized as a light yellow solid.

[0309] Step 2:

[0310] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 26-1 (102 mg) and piperidine (0.1 mL) were used as raw materials to synthesize the target product V-26 (35 mg). MS (ESI) m / z: 1121.3 [M+H] +. HRMS (ESI): Anal. Calcd for C 57 H 93 N4O 18 [M+H]+:1121.6441.Found:1121.6438.

[0311] Example 27 Synthesis of Compound V-27

[0312]

[0313] Step 1:

[0314] With reference to the synthesis method of step 2 in Example 1, 18-2 (100 mg, 0.083 mmol, 1 eq), PyBOP (52 mg, 0.1 mmol, 1.2 eq), 27-1 (40 mg, 0.125 mmol, 1.5 eq, ACS Infect Dis. 2020, 14, 2029-2044) and DIPEA (49 μL, 0.291 mmol, 3.5 eq) were added sequentially to a single-necked bottle containing DMF (3 mL). The outer wall of the single-necked bottle was wrapped with aluminum foil and protected by argon. The reaction was stirred at room temperature overnight and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (60 mL), filtered to obtain the crude product of 27-2, and purified by silica gel column chromatography (eluent: dichloromethane / methanol = 10: 1 to 5: 1) to obtain compound 27-2 (65 mg) as a light yellow solid.

[0315] Step 2:

[0316] Referring to the synthesis method of Step 3 in Example 1, 27-2 (65 mg) and piperidine (0.1 mL) were added sequentially to a single-necked flask containing DMSO (3 mL). The outer wall of the single-necked flask was wrapped with aluminum foil and protected by argon. The reaction was stirred at room temperature for 2 hours, and the reaction progress was monitored by HPLC. After the reaction was complete, the reaction solution was poured into vigorously stirred methyl tert-butyl ether (100 mL) and filtered. The resulting solid was washed with methyl tert-butyl ether (40 mL x 2), filtered, and dried in vacuo to obtain the target product V-27 (29 mg) as a light yellow solid. 1 H NMR (600MHz, DMSO-d6) δ8.25(br,1H),8.12–7.94(m,1H),6.50–6.02(m,12H),5.93(dd,J=15.3,9.0Hz,2H),5.43(dd,J=14.9,10.2Hz,1H),5. 28–5.15(m,1H),5.03–4.49(m,8H),4.42–4.29(m,4H),4.28–4.14(m,4 H),4.13–3.93(m,6H),3.74–3.65(m,2H),3.63(s,2H),3.56–3.49(m,2 H),3.46(s,1H),3.45(s,1H),3.22–3.09(m,4H),2.88–2.73(m,1H),2. 36–2.22(m,1H),2.16(d,J=6.1Hz,2H),1.98–1.88(m,2H),1.87(s,1H) ,1.75–1.68(m,1H),1.62–1.35(m,8H),1.34–1.21(m,3H),1.18–1.14( m,3H),1.12–1.10(m,3H),1.03(d,J=6.1Hz,3H),0.91(d,J=7.0Hz,3H). 13C NMR(151MHz,DMSO-d6)δ172.71,170.60(2C),136.83,136.29,133.86,133.72,133.34,133.22,132.56(2C ),132.40,132.27,132.11,131.92,131.24,129.00,97.15,97.05,77.20,75.18,73.80,73.59,73.34,69. 98,69.20,69.04,68.88,67.77,66.24,65.24,64.79,57.20,54.76,46.23,44.79,44.45,43.72,43.04,42 .51(2C),42.06,40.06,37.75,37.00,35.10,29.04,18.56,18.12,17.01,12.12.MS(ESI)m / z:1023.1[M+H] + .HRMS(ESI):Anal.Calcd for C 51 H 83 N4O 17 [M+H] + :1023.5748,Found:1023.5776.

[0317] Example 28 Synthesis of Compound V-28

[0318]

[0319] Step 1:

[0320] Referring to the synthesis method of step 1 in Example 27, using DMF as solvent, 20-2 (100 mg, 0.082 mmol, 1 eq), PyBOP (52 mg, 0.099 mmol, 1.2 eq), 27-1 (40 mg, 0.123 mmol, 1.5 eq, ACS Infect Dis. 2020, 14, 2029-2044) and DIPEA (49 μL, 0.288 mmol, 3.5 eq) as raw materials, compound 28-1 (55 mg) was synthesized as a light yellow solid.

[0321] Step 2:

[0322] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 28-1 (55 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-28 (21 mg) as a light yellow solid. 1H NMR(600MHz,DMSO-d6)δ8.10–7.99(m,1H),7.94–7.84(m,1H),6.55–5.82(m,12H),5.50–5.37(m,1H),5.27–5.14(m,1H ),4.94–4.52(m,4H),4.41–4.30(m,2H),4.28–4.16(m,2H),4.13–3.97(m,4H),3.24–3.01(m,8H),2.93–2.78(m,2H),2 .74–2.58(m,2H),2.42–2.21(m,3H),2.17–2.14(m,2H),2.01–1.85(m,2H),1.84–1.81(m,2H),1.76–1.66(m,1H),1.64 –1.21(m,11H),1.16–1.13(m,3H),1.12–1.09(m,3H),1.05–1.02(m,3H),0.97–0.85(m,3H).MS(ESI)m / z:1059.4[M+Na] + .HRMS(ESI):Anal.Calcd for C 52 H 85 N4O 17 [M+H] + :1037.5904,Found:1037.5924.

[0323] Example 29 Synthesis of Compound V-29

[0324]

[0325] Step 1:

[0326] Referring to the synthesis method of step 1 in Example 27, using DMF as solvent, 20-2 (100 mg, 0.082 mmol, 1 eq), PyBOP (52 mg, 0.099 mmol, 1.2 eq), 27-1 (40 mg, 0.123 mmol, 1.5 eq, ACS Infect Dis. 2020, 14, 2029-2044) and DIPEA (49 μL, 0.288 mmol, 3.5 eq) as raw materials, compound 29-1 (47 mg) was synthesized as a light yellow solid.

[0327] Step 2:

[0328] Referring to the synthesis method of step 3 in Example 1, using DMSO as solvent, 29-1 (47 mg) and piperidine (0.1 mL) as raw materials, the target product V-29 (21 mg) was synthesized as a light yellow solid. 1H NMR(600MHz,DMSO-d6)δ7.99(br,1H),7.64(d,J=9.0Hz,1H),6.51–6.01(m,12H),5.97–5.89(m,1H),5.85(br,1H),5.43(dd,J=14.9,10.0Hz,2H),5.27–5.17(m,1H),4.99–4.53(m,4H),4.39–4.32(m,2H),4.27–4.18(m,2H),4.10–4.04(m,1H),4.04–4.00(m,1H),3.69(t,J=9.3Hz,2H),3.61–3.56(s,2H),3.55–3.49(m,2H),3.48–3.43(m,2H),3.20–3.09(m,4H),2.63(dt,J=12.5,6.2Hz,1H),2.27(s,3H),2.26(s,1H),2.16(d,J=6.1Hz,2H),2.00–1.80(m,3H),1.75–1.68(m,1H),1.64–1.44(m,5H),1.43–1.35(m,2H),1.31(d,J=13.5Hz,2H),1.28–1.21(m,2H),1.15(d,J=6.0Hz,3H),1.12–1.11(m,3H),1.03(d,J=6.2Hz,3H),0.91(d,J=7.0Hz,3H). 13 C NMR(151MHz,DMSO-d6)δ172.17,170.85,170.60,136.80,136.60,133.89,133.72,133.42,133.23,132.53(2C),132.24(2C),131.92(2C),131.25,128.92,97.12(2C),77.21,75.22,73.82,73.59,73.29,70.41,69.24(2C),68.88,67.77,66.25,65.40,64.69,57.12,54.36,54.06,46.30,44.78,44.58,42.49,42.05,41.16,41.00,40.43,40.06,36.95,36.12,35.12,29.07,18.55,18.10,17.01,12.11.MS(ESI)m / z:1037.0[M+H] + .HRMS(ESI):Anal.Calcdfor C 52 H 85 N4O 17[M+H] + :1037.5904,Found:1037.5926.

[0329] Example 30 Synthesis of Compound V-30

[0330]

[0331] Compound V-29 (30 mg, 0.029 mmol, 1 eq) was added to a solution of mandelic acid (9 mg, 0.058 mmol, 2 eq) in water (3 mL). The mixture was stirred at room temperature for 3 min, filtered, and the filtrate was lyophilized under vacuum to give the target product V-30 (38 mg) as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ8.42(s,1H),8.30(d,J=8.4Hz,1H),7.39(d,J=7.5Hz,4H),7.26(t,J=7. 5Hz,4H),7.19(t,J=7.3Hz,2H),6.51–6.02(m,12H),5.97–5.83(m,2H),5.44(dd,J=14.9,9.9Hz ,2H),5.21(d,J=7.1Hz,1H),4.71(s,5H),4.35(s,2H),4.31(s,2H),4.23(t,J=9.4Hz,3H),4.10 –3.99(m,4H),3.70(s,1H),3.65(s,1H),3.57–3.49(m,2H),3.49–3.40(m,4H),3.38(d,J=5.2Hz, 1H),3.35(d,J=5.2Hz,1H),3.30(d,J=5.7Hz,1H),3.28(d,J=5.9Hz,1H),3.17(t,J=7.5Hz,2H), 3.09(d,J=8.0Hz,2H),2.92–2.78(m,2H),2.54(s,1H),2.41(s,3H),2.28(q,J=7.6,7.2Hz,2H),2 .16(d,J=6.2Hz,2H),1.95–1.83(m,3H),1.72(d,J=7.8Hz,1H),1.62–1.49(m,4H),1.48–1.21(m ,9H),1.17(d,J=5.2Hz,3H),1.11(d,J=6.1Hz,3H),1.04(d,J=6.1Hz,3H),0.91(d,J=7.0Hz,3H). 13CNMR(126MHz,DMSO)δ174.89(2C),172.84,170.59,167.33,142.55(2C),136.84,136.25,133.83,133.69,133.29,133.22,132. 55(2C),132.39,132.26,132.10,131.90,131.22,129.03,127.62(4C),126.59(2C),126.43(4C),97.14,97.00,75.12,73.82,7 3.59,73.38,73.25(2C),72.51(2C),69.63,69.22,68.91,67.78,66.24,65.26,64.77,63.09,57.73,54.99,51.24,46.35,44.7 6,44.41,42.51,42.06,40.43,38.87,37.01,36.83,35.09,33.85,29.05,18.52,18.09,16.99,12.09.MS(ESI)m / z:1037.2[M+H] + .HRMS(ESI):Anal.Calcd for C 52 H 85 N4O 17 [M+H] + :1037.5909,Found:1037.5904.

[0332] Example 31 Synthesis of Compound V-31

[0333]

[0334] Referring to the synthesis method in Example 30, compound V-29 (100 mg, 0.097 mmol, 1 eq) was added to a solution of L-glutamic acid (29 mg, 0.193 mmol, 2 eq) in water (10 mL). The mixture was stirred at room temperature for 5 min, filtered, and the filtrate was lyophilized under vacuum to obtain the target product V-31 (118 mg) as a yellow solid. 1H NMR(500MHz,Deuterium Oxide)δ6.78–5.91(m,13H),5.38(s,1H),5.26(s,1H),4.84(s,2H),4.74(s,2H),4.60(s,1H),4.52(s,1H),4.43(s,1H), 4.38(s,1H),4.28(s,2H),4.05–3.92(m,4H),3.80(dd,J=7.0,5.1Hz,4H),3.52–3.38(m,3H),3.30–3.16(m,3H),2.87–2. 80(m,4H),2.75(s,1H),2.46(d,J=3.3Hz,2H),2.44(d,J=3.0Hz,2H),2.43(d,J=2.6Hz,2H),2.25–2.06(m,9H),1.96–1.6 6(m,6H),1.55(s,2H),1.35(d,J=5.3Hz,3H),1.26(d,J=5.7Hz,3H),1.18(s,3H),1.05(s,3H).MS(ESI)m / z:1037.2[M+H] + .HRMS(ESI):Anal.Calcd for C 52 H 85 N4O 17 [M+H] + :1037.5900,Found:1037.5904.

[0335] Example 32 Synthesis of Compound V-32

[0336]

[0337] Referring to the synthesis method in Example 30, using water as solvent, compound V-29 (30 mg, 0.029 mmol, 1 eq) and DL-aspartic acid (8 mg, 0.058 mmol, 2 eq) as raw materials, the target product V-32 (37 mg) was synthesized as a yellow solid. 1HNMR(500MHz,Deuterium Oxide)δ6.76–6.00(m,13H),5.47–5.20(m,1H),4.84(s,2H),4.60(s,1H) ),4.55–4.46(m,1H),4.43(s,2H),4.33–4.22(m,2H),4.00(s,2H),3.95( dd,J=8.4,3.9Hz,4H),3.80(td,J=6.5,3.2Hz,1H),3.69(d,J=4.4Hz,1H),3.67(d,J=4.4Hz,1H),3.60(d,J=6.5Hz,1H),3.57(d,J=6.5Hz,1H),3.5 1–3.41(m,3H),3.28–3.16(m,3H),2.88(d,J=3.9Hz,1H),2.86–2.81(m,5 H),2.79–2.70(m,4H),2.53–2.34(m,1H),2.26–2.04(m,3H),1.96–1.66( m,5H),1.55(s,3H),1.37–1.33(m,3H),1.28–1.24(m,3H),1.21(t,J=7.2 Hz,2H),1.19–1.15(m,3H),1.07–1.03(m,3H).MS(ESI)m / z:1037.2[M+H] + .HRMS(ESI):Anal.Calcd for C 52 H 85 N4O 17 [M+H] + :1037.5896,Found:1037.5904.

[0338] Example 33 Synthesis of Compound V-33

[0339]

[0340] Step 1:

[0341] Referring to the synthesis method of step 1 in Example 27, using DMF as solvent, 9-2 (100 mg, 0.083 mmol, 1 eq), PyBOP (52 mg, 0.1 mmol, 1.2 eq), 33-1 (39 mg, 0.125 mmol, 1.5 eq, Synlett. 2004, 3, 453-456) and DIPEA (28 μL, 0.166 mmol, 2 eq) as raw materials, compound 33-2 (54 mg) was synthesized as a light yellow solid.

[0342] Step 2:

[0343] Referring to the synthesis method of step 3 in Example 1, DMSO was used as solvent, 33-2 (54 mg) and piperidine (0.1 mL) as raw materials to synthesize the target product V-33 (32 mg) as a light yellow solid. MS (ESI) m / z: 1062.5 [M+H] + .HRMS(ESI):Anal.Calcd for C 54 H 87 N4O 17 [M+H] + :1062.6032,Found:1062.6054.

[0344] Test Example 1 In vitro antifungal activity test of preferred compounds

[0345] 1.1 Methods: The minimum inhibitory concentration (MIC) of the test samples against the test strains was determined using the agar two-fold dilution method recommended by the Clinical and Laboratory Standards Institute (CLSI) antimicrobial susceptibility testing procedure [M44-A2, 2013].

[0346] Culture medium and conditions:

[0347] Culture medium: Sabouraud agar

[0348] Culture conditions: Incubate at 35-37°C for 48 hours and observe the results.

[0349] Table 1. Clinical isolates tested for in vitro antimicrobial activity screening

[0350]

[0351]

[0352] 1.2 Experimental plan:

[0353] The minimum inhibitory concentration (MIC) of the compounds against fungi was determined by the agar double dilution method recommended by the Clinical and Laboratory Standards Institute (CLSI) [Method for Antifungal Disk Diffusion Susceptibility Testing of Yeast; Approved Guideline-Second Edition (Vol, 29, No. 17); M44-A2]. Concentration (MIC); Specific method: add 1 ml of drug solution to a sterile plate, then add 14 ml of melted 50°C Sabouraud agar medium and mix well to make the final drug concentration in each plate 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125, 0.06, 0.03, 0.015, 0.008 mg / L in sequence; after cooling, use a multi-point inoculator (MIT-P, SUKUMA) to inoculate fungi with an inoculum of about 10 4 CFU / ml, cover the dish. Culture in a 35-37℃ incubator. Observe and record the results after 48 hours of culture. The lowest concentration of the drug at which no bacteria grow in the dish is determined as the minimum inhibitory concentration (MIC). The quality control compound is amphotericin B for injection, and the quality control strain is Candida krusei ATCC 6258. The MIC reference value standard refers to CLSI2016 (Reference Method for AntifungalDisk Diffusion Susceptibility Testing of Yeasts; Fourth InformationalSupplement; M27-S4). The MIC reference range of amphotericin B for injection against standard quality control strains is shown in Table 2 below.

[0354] Table 2 MIC range of amphotericin B against standard quality control bacteria (CLSI2012 judgment criteria)

[0355]

[0356] 1.3 In vitro antifungal test results of the compounds of the present invention

[0357] The results are shown in Table 3:

[0358] Table 3 In vitro antifungal activity of diamide-modified amphotericin B derivatives (MIC, μg / mL)

[0359]

[0360]

[0361] Table 3 In vitro antifungal activity of diamide-modified amphotericin B derivatives (MIC, μg / mL)

[0362]

[0363] Table 3 In vitro antifungal activity of diamide-modified amphotericin B derivatives (MIC, μg / mL)

[0364]

[0365]

[0366] Table 3 In vitro antifungal activity of diamide-modified amphotericin B derivatives (MIC, μg / mL)

[0367]

[0368] As shown in Table 3, compounds V-(1-9) derivatized with acylhydrazides at the carboxyl terminus exhibited superior activity against Candida albicans, Candida krusei, and Neococcus neoformans. Compound V-2 showed the strongest antibacterial activity in the series, significantly outperforming amphotericin B. Most of the diamide-substituted amphotericin B derivatives V-(10-29) exhibited stronger in vitro antibacterial activity than amphotericin B against various fungi. In particular, compounds V-12, V-13, V-17, V-18, V-20, V-22, V-25, V-26, V-27, V-28, and V-29 exhibited significantly superior comprehensive antibacterial activity to amphotericin B. V-29 exhibited 62-250-fold greater activity against Candida albicans, 4-8-fold greater activity against Candida krusei, 2-16-fold greater activity against Candida parapsilosis, and 16-32-fold greater activity against Candida tropicalis compared to amphotericin B.

[0369] In conclusion, the compounds of the present invention have excellent in vitro antibacterial activity, and the in vitro antibacterial activity of the representative compounds is significantly better than that of amphotericin B.

[0370] Test Example 2 In vitro hemolytic activity test of the compounds of the present invention

[0371] 2.1 Experimental plan:

[0372] The cells used in this experiment were human red blood cells, which were prepared into a suspension (concentration of 4 x 10 8cells / mL) for use. All test compounds were dissolved in dimethyl sulfoxide for use. 736 μL of PBS solution and 14 μL of dimethyl sulfoxide solution of the test compound were added to a 1.5 mL PCR tube, and finally 750 μL of human red blood cell suspension was added. In the final solution, the solubility of the test compound was 200, 150, 100, 80, 50, 30, 20, 10, 5, 1 μM (the highest drug concentration was selected according to the actual situation). For each experiment, a blank control experiment (only dimethyl sulfoxide, no test compound) and a positive control experiment (addition of 1% Triton X100) were carried out. Then, the PCR tube was incubated at 37°C for 1 hour and then centrifuged at 4°C (1500g) for 5 minutes. The supernatant was taken and their optical density (OD 560 The calculation formula of the cleavage rate of a compound at a specific concentration is as follows:

[0373]

[0374] The positive control experimental group was calculated as 100% human red blood cell lysis. Finally, GraphPad Prism 6 was used to calculate the concentration of each compound that lysed 50% of human red blood cells and calculated as EC 50 (μM).

[0375] 2.2 In vitro hemolytic activity test results of the compounds of the present invention

[0376] Table 4 In vitro hemolytic activity of some amphotericin B derivatives

[0377]

[0378] As can be seen from the table above, the compounds of the present invention exhibit significantly improved in vitro hemolytic activity compared to amphotericin B. In particular, compounds V-15 and V-29 exhibited 19-fold and 56-fold lower hemolytic activity, respectively, compared to amphotericin B. This indicates that the compounds of the present invention exhibit lower hemolytic toxicity and significantly improved safety compared to amphotericin B.

[0379] Combined with the in vitro antibacterial activity results in Table 3 above, it is fully demonstrated that the compound of the present invention has a larger therapeutic window and a higher efficacy index than amphotericin B.

[0380] Test Example 3 Pharmacokinetics of the Compounds of the Invention in Mice

[0381] 3.1 Experimental plan:

[0382] In this experiment, healthy male CD-1 mice weighing 18-22 g (from the Shanghai Laboratory Animal Center, Chinese Academy of Sciences) were selected and intravenously injected with the test compound and amphotericin B. The dosing regimen is shown in Table 5 below:

[0383] Table 5 Dosage regimen of the test compounds

[0384]

[0385]

[0386] Animals were fasted for 12 hours before the experiment, with free access to water. Two hours after drug administration (drug dissolved in DMSO:PEG300:EtOH:NaCl = 5:40:5:50, v / v / v), a single meal was resumed. Approximately 25 μL of blood was collected from each animal via the femoral vein and placed in a heparinized tube. The blood sample was placed on ice and centrifuged within 30 minutes to separate plasma (5000 rpm, 10 minutes, 4°C). Blood was collected at 0.05, 0.25, 0.75, 2, 4, 8, and 24 hours after administration. Collected plasma was stored at -20°C.

[0387] 3.2 Results of pharmacokinetic studies of the compounds of the present invention in mice

[0388] The pharmacokinetic parameters of compounds V-15, V-27, and V-29 and amphotericin B administered intravenously to mice are shown in Table 6.

[0389] Table 6 Main pharmacokinetic parameters of V-15, V-27, V-29, and amphotericin B injected intravenously into mice (1 mg / kg, N=3)

[0390]

[0391] As shown in Table 6, after intravenous administration, representative compounds V-15, V-27, and V-29 of the present invention reduced clearance by 76%, 77%, and 87%, respectively, compared to amphotericin B. These clearances reached 0.95, 0.91, and 0.53 mL / min / kg, respectively, resulting in 4.3-, 4.6-, and 7.1-fold increases in drug exposure. V-29, in particular, exhibited the best metabolic properties, significantly outperforming amphotericin B. These data demonstrate that the compounds of the present invention exhibit superior metabolic properties and improved drugability, likely due to the diamide substitution in the compounds, which disrupts the amphiphilic nature of the amphotericin B molecule and reduces its ability to form intermolecular aggregates, making it more likely to bind to ergosterol for antimicrobial activity rather than cholesterol for toxicity.

[0392] Test Example 4 In vivo antifungal activity test of the compounds of the present invention

[0393] 4.1 Experimental plan:

[0394] In this experiment, healthy KM mice weighing 18-22g were selected. After quarantine, they were randomly divided into groups and given free food and water. The infection source was Candida albicans (strain 20-1). The efficacy of a single intravenous injection of compound V-31 and the control drug Amphamide glutamate (ACS Infect. Dis. 2020, 6, 2029-2044) on mice infected with Candida albicans was tested. The test drug was prepared with normal saline and diluted to the required concentration solution, which was prepared and used immediately.

[0395] 4.1.1 Preparation of bacterial solution

[0396] Select several colonies from a 24-hour-old Sabouraud agar plate and prepare a bacterial suspension directly in sterile saline. Adjust the concentration of the suspension to a McFarland turbidimetric value of 0.5-2.0. Dilute the adjusted suspension with saline and use immediately.

[0397] 4.1.2 Determination of minimum lethal dose

[0398] Healthy mice were randomly divided into groups of 5, half male and half female. Four days before infection and one day before infection, respectively, mice were immunosuppressed by intraperitoneal injection of 150 mg / kg and 100 mg / kg of cyclophosphamide. On the day of infection, 0.5 ml of the bacterial solution described above at different dilutions was injected into the tail vein of each mouse. The mice were observed for 7-14 days after infection, and the number of mice that died was recorded. The minimum lethal dose (MLD) that caused 100% mortality in the mice was used as the infection dose for in vivo experiments.

[0399] 4.1.3 In vivo antifungal activity assay

[0400] The mice were deprived of food and water 18 hours before the experiment and randomly divided into 8 mice per group, half male and half female. 150 and 100 mg / kg cyclophosphamide were injected intraperitoneally on the 4th and 1st day before inoculation, respectively. On the day of infection, the minimum lethal bacterial liquid was drawn and injected into the mice through the tail vein, 0.5 ml per mouse, to establish a mouse model of systemic infection caused by Candida albicans. One hour after the animals in each treatment group were infected with the bacterial liquid, the drug was injected into the tail vein in a single dose according to the designed dose. One hour after the infection, the animals in the infection control group were injected into the tail vein with an equal volume of normal saline. 7-14 days after administration, the survival of the animals was observed and recorded every day. According to the number of mouse deaths, the half effective dose (ED) was calculated according to the Bliss method. 50 and 95% confidence limits, and statistical analysis was performed.

[0401] 4.2 Results of the antifungal activity test of the compounds of the present invention in mice

[0402] The in vivo antifungal activities of compound V-33 and Amphamide glutamate against Candida albicans-infected mice (single intravenous injection, N=8) are shown in Table 7.

[0403] Table 7 In vivo antifungal activity of compound V-33 and Amphamide glutamate against Candida albicans infected mice (single intravenous injection, N=8)

[0404]

[0405] As shown in Table 7, the representative compound V-33 of the present invention has an antifungal activity against Candida albicans-infected mice increased by about 1.5 times compared with the control drug Amphamide glutamate when administered intravenously once, and has an antifungal activity against Candida albicans-infected mice increased by about 1.5 times compared with amphotericin B (ED 50 =1.08 mg / kg, ACS Infect. Dis. 2020, 6, 2029-2044), the antifungal activity in vivo was increased by about 4 times.

[0406] Combined with the results of hemolytic activity experiments and in vivo pharmacokinetic tests, the representative compound V-29 of the present invention has significantly reduced toxicity and significantly improved activity compared with amphotericin B, can greatly expand the therapeutic window, and exhibits better pharmacokinetic properties, with obvious drug-making advantages.

[0407] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound of formula V, or a pharmaceutically acceptable salt thereof: in, R1 are each independently selected from the following group: -NR4-(L) p -Ra, -NR4(CH2) n R5, -NHCH2CH2NH2, wherein Ra is selected from the group consisting of H, C1-C6 alkyl, C1-C6 alkoxy, -NH2, 6-10 membered aryl, 5-10 membered heteroaryl, and -COR3; L is selected from the group consisting of NH, NHC(O); R4 are each independently selected from the group consisting of H, C1-C6 alkyl; R5 is selected from the group consisting of H, substituted or unsubstituted C1-C6 alkyl; R6 is selected from the group consisting of H, OH, C1-C6 alkyl, and 5-12 membered heterocyclic group; R2 are each independently selected from the following group: H, R3 is NH2; p is selected from the group consisting of 0, 1 or 2; n is selected from the group consisting of 0, 1, 2, 3, 4, or 5; The term "substituted" refers to the replacement of one or more hydrogen atoms on a group with a hydroxy group.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein The compound is selected from the following table:

3. The method for preparing the compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: Including steps: In an inert solvent, the protecting group is removed with compound M-2 to obtain a compound of formula V; Wherein, the PG is an amino protecting group; the R2' is selected from the following group: chemical bond, -C(O)(L) p -Rc'; wherein Rc' is a group formed when Rc loses a hydrogen atom; and when R2' is a chemical bond, R1 is -NR4-(L) p -Ra, and at least one L in the R1 group is NH or NHC(O); wherein Rc is independently selected from the following group: (CH2)nNHR4, NHR4, or a substituted or unsubstituted 3-8 nitrogen-containing heterocyclic group; The remaining groups are as defined in claim 1.

4. The method for preparing the compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein: Including steps: In an inert solvent, the protecting group is removed with compound M-4 to obtain a compound of formula V; Wherein, the PG is an amino protecting group; The R1' is a group formed by R1 losing a H atom; The R2' is selected from the following group: chemical bond, -C(O)(L) p -Rc'; wherein Rc' is a group formed when Rc loses a hydrogen atom; and when R2' is a chemical bond, R1 is -NR4-(L) p -Ra, and at least one L in the R1 group is NH or NHC(O); wherein Rc is independently selected from the following group: (CH2)nNHR4, NHR4, or a substituted or unsubstituted 3-8 nitrogen-containing heterocyclic group; The remaining groups are as defined in claim 1.

5. A pharmaceutical composition, characterized in that Contains the following components: 1) a therapeutically effective amount of one or more compounds according to claim 1, or a pharmaceutically acceptable salt thereof; and 2) Pharmaceutically acceptable carriers or excipients.

6. Use of the compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that: Used for preparing a pharmaceutical composition for preventing or treating infectious diseases caused by fungal infection.

7. The use according to claim 6, characterized in that The fungus is a dermatophyte.

8. The use according to claim 6, characterized in that The fungus is selected from the group consisting of Candida, Cryptococcus neoformans, Aspergillus, Combinatorial fungi, or a combination thereof.

9. The use according to claim 8, characterized in that The Aspergillus is selected from the group consisting of Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Aspergillus niger, or a combination thereof.

10. An intermediate for preparing the compound of formula V according to claim 1, characterized in that: The intermediate is selected from the group consisting of: Wherein, the PG is an amino protecting group, and the amino group N connected thereto constitutes a carbamate, an amide, an N-alkylamine or an N-arylamine; The R1' is a group formed by R1 losing a H atom; R2' is selected from the following group: chemical bond, -C(O)(L) p -Rc'; wherein Rc' is a group formed when Rc loses a hydrogen atom; and when R2' is a chemical bond, R1 is -NR4-(L) p -Ra, and at least one L in the R1 group is NH or NHC(O); wherein Rc is independently selected from the following group: (CH2)nNHR4, NHR4, or a substituted or unsubstituted 3-8 nitrogen-containing heterocyclic group; the definitions of the remaining groups are as described in claim 1.

11. An intermediate for preparing the compound of formula V according to claim 10, characterized in that: The protecting group is selected from the group consisting of Fmoc, Boc, Cbz, Alloc, Teoc, methoxycarbonyl or ethoxycarbonyl, Tfa, Pht, Tos, Ns, pivaloyl, benzoyl, Bn, PMB, Trt, and Dmb.

12. An intermediate for preparing the compound of formula V according to claim 10, characterized in that: The protecting group is selected from the group consisting of Fmoc, Boc, Tos, and Cbz.

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