CN114249721A - Vinblastine PEG derivative and application thereof in preparation of drugs for treating diabetic peripheral neuropathy, diabetic foot and pulmonary fibrosis - Google Patents

Abstract

The invention relates to a PEGylated vincamine derivative and application thereof. The invention particularly discloses a PEGylated vincamine derivative shown as a formula I and a formula II, and pharmaceutically acceptable salts, metabolic precursors, metabolic products or isomers thereof of the PEGylated vincamine derivative. Experiments show that the vincamine derivative can promote the growth of peripheral sensory neuron protrusion, improve the nerve conduction speed and sensory loss symptoms of the diabetic rat, and promote the healing of foot ulcer wounds of the diabetic rat, so that the vincamine derivative has a good treatment effect on diabetic complication peripheral neuropathy and diabetic feet. Experiments show that the vincamine derivatives can also obviously reduce the pulmonary fibrosis degree of mice induced by bleomycin and protect pulmonary tissues.

Description

Vinblastine PEG derivative and application thereof in preparation of drugs for treating diabetic peripheral neuropathy, diabetic foot and pulmonary fibrosis

Technical Field

The invention belongs to the field of medicine and chemical synthesis, and particularly relates to a preparation method of a vincamine derivative, a pharmaceutical composition and application of the vincamine derivative in treating peripheral neuron axon injury, peripheral related neuropathy, diabetic foot and pulmonary fibrosis.

Background

Diabetes is a serious worldwide disease. It is estimated that by 2045 years, the population of diabetics will reach 6.29 billion. Diabetes has now become a third non-infectious disease that threatens human health and life, following cardiovascular disease and tumors. In China, diabetes mellitus has changed from rare disease to epidemic disease, and the prevalence rate rises dramatically from 0.67 percent to 11.6 percent in average in nearly 30 years. If the diabetes patient does not adopt an effective control treatment method, the diabetes patient is easily troubled by various diseases complicated by diabetes, including diabetic neuropathy, nephropathy, diabetic foot and the like, and binocular blindness, renal failure and even limb disability and death are caused.

Diabetic Peripheral Neuropathy (DPN) is a chronic neuropathy caused by long-term hyperglycemia, has a disease rate of 50% to 80%, and is one of the most common chronic complications of diabetes. Diabetic peripheral neuropathy can involve the motor, sensory and autonomic nerves, resulting in movement and sensory disorders. Clinically, the vibration and heat perception thresholds of diabetic peripheral neuropathy patients increase, and these thresholds progress to sensory loss as all fibers in the peripheral nerve denature. Currently, the diagnostic methods for diabetic peripheral neuropathy mainly include sensory tests, electrophysiological tests, nerve fiber morphological tests, imaging tests, and the like. However, the clinical manifestations of diabetic peripheral neuropathy are various, the classification methods are different at home and abroad, the screening and examination methods are various, and the diagnosis standards are different, so that the prevention and treatment of the diabetic peripheral neuropathy are always clinical difficulties. Although scientists have breakthrough progress on the research of diabetic peripheral neuropathy with the rapid development of scientific technology, specific medicines for radically treating diabetic peripheral neuropathy still lack. Therefore, research and development of new therapeutic strategies and new compounds is slow.

Diabetic Foot (DF) is a disease in which Diabetic vasculopathy and/or neuropathy and infection are caused by long-term hyperglycemia, and then the tissues of the foot or lower limb of a Diabetic patient are necrotized. The diabetic foot is one of the most serious complications of diabetes, is one of the main causes of disability and death of the diabetic patient, and is also a major public health problem causing heavy social burden. With the increasing incidence of diabetes, the number of patients with diabetic foot has also increased year by year. The incidence rate of diabetic foot of diabetic patients over 50 years old in China is up to 8.1%. Unfortunately, the prognosis of diabetic foot is poor and even higher than the mortality and disability rates of most cancers (except lung cancer, pancreatic cancer, etc.). And the diabetic foot treatment cost is high, the treatment needs to be accepted for a long time, the foot ulcer of a patient is healed mainly through medical care in clinic, and the danger of amputation is avoided. However, these treatments do not completely cure the diabetic foot and are prone to relapse. Therefore, the research on the pathogenesis of the diabetic foot and the development of novel effective therapeutic drugs for the diabetic foot have very important practical significance.

Pulmonary Fibrosis (PF) is a chronic, progressive, lethal lung disease, the terminal stage of many diseases, and the pathogenesis of the disease remains unclear. Pulmonary fibrosis seriously affects the respiratory function of the human body, manifested as dry cough, progressive dyspnea (insufficient subjective qi), and the respiratory function of the patient is continuously worsened with the aggravation of the disease condition and the lung injury. The incidence and mortality of idiopathic pulmonary fibrosis increases year by year, with an average survival period of only 2.8 years after diagnosis, with a mortality rate higher than that of most tumors, known as a "neoplastic-like disease". At present, pirfenidone, nintedanib, oxygen therapy, invasive/non-invasive mechanical ventilation and lung transplantation are mostly adopted for treating patients with pulmonary fibrosis. However, these methods have high cost, insignificant curative effect, and limited wide application due to insufficient transplant donor or immunological rejection and poor medical compliance of patients. Therefore, it is very important to actively explore the pathogenesis of pulmonary fibrosis and to develop new intervention drugs.

Vincamine (Vincamine) is a monoterpene indole alkaloid found in Madagas Elephantopus. It can increase cerebral blood flow, oxygen consumption and glucose utilization rate, and improve dementia and dysmnesia. Vincamine has been marketed in the united states as a health product with nootropic effects. However, like many natural products, vincamine also suffers from poor water solubility, unstable metabolism, short duration of action, and the like. From the previous work, vincamine has potential in treating diabetic complications and pulmonary fibrosis. Therefore, the search for novel vincamine derivatives is of great significance for the treatment of diabetic complications and pulmonary fibrosis.

Polyethylene glycol (PEG) is a water-soluble polyether of relatively low molecular weight obtained by the stepwise addition polymerization of ethylene oxide with water or ethylene glycol. The low molecular weight poly-ethylene glycol is colorless, odorless and hygroscopic viscous liquid, has ether chains and hydroxyl groups in molecules, has unique solubility and good biocompatibility, and has important application prospect in the fields of medicines, materials, engineering and the like.

Disclosure of Invention

The technical problem to be solved by the invention is to utilize the characteristics of no toxicity and easy combination of PEG, connect the PEG with the vincamine and overcome the characteristic of the vincamine that the vincamine is insoluble in water. The invention provides a vincamine derivative, a preparation method thereof, a pharmaceutical composition and application. The PEGylated vincamine derivative has stable structure and good water solubility, and has good treatment effect on diabetic peripheral neuropathy, diabetic foot and pulmonary fibrosis.

The invention solves the technical problems through the following technical scheme.

The invention provides a vincamine PEG derivative, which comprises compounds shown as a formula I and a formula II, pharmaceutically acceptable salts, metabolic precursors, metabolites or isomers thereof, and has the following structure:

in formula I and formula II:

l is a single bond,

Amino acids, or, di-hexapeptides formed by amino acid condensation;

x and Y are independently selected from single bonds、

Amino acids, or, di-hexapeptides formed by amino acid condensation;

t is an integer of 1 to 6;

R¹is hydrogen, unsubstituted or R^1-1Substituted C_1-6Alkyl radical, C_6-10Aryl radical, C_3-10Cycloalkyl, one or more than one heteroatom selected from N, O, S, "5-10 membered heteroaryl with 1-3 heteroatoms", or "4-10 membered heterocycloalkyl with 1-3 heteroatoms" selected from N, O, S, C_3-10Cycloalkyl- (C)_1-4Alkyl) -, or, C_6-12Aryl radical- (C)_1-4Alkyl) -;

R^1-1is halogen, hydroxy, amino or C_1-4An alkyl group;

n is an integer of 0 to 500;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-polyethylene glycol of linear, dendritic, star-shaped or hyperbranched structure;

r is-O (R)²)、-NH(R³) Or, -COOH;

R²is hydrogen or C_1-6An alkyl group;

R³is hydrogen or C_1-6An alkyl group.

When L is an amino acid, the amino acid is alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, or histidine;

when L is a di-hexapeptide formed by condensing amino acids, the amino acids are the same or different;

when L is a di-hexapeptide formed by condensing amino acids, the di-hexapeptide is a di-tetrapeptide;

when X and Y are independently selected from amino acids, said amino acids are alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, or histidine;

when X and Y are independently selected from the group consisting of di-hexapeptides formed by the condensation of amino acids, said amino acids are the same or different;

when X and Y are independently selected from the group consisting of di-hexapeptides formed by the condensation of amino acids, said di-hexapeptides are di-tetrapeptides;

when R is¹Is unsubstituted or R^1-1Substituted C_1-6When it is alkyl, said R^1-1Is one or more, when there are more than one R^1-1When R is said^1-1May be the same or different;

when R is¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

when R is¹Is C_6-10When aryl, said C_6-10Aryl is phenyl;

when R is¹Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl being C_3-6A cycloalkyl group;

when R is¹Is C_6-12Aryl radical- (C)_1-4Alkyl) -said C_6-12Aryl radical- (C)_1-4Alkyl) -is phenyl- (C)_1-2Alkyl) -;

n is an integer of 0 to 300;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-a linear polyethylene glycol;

when R is²Is C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

when R is³Is C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group.

When L is an amino acid, the amino acid is alanine, phenylalanine, methionine, glutamic acid, aspartic acid, valine, leucine, and proline, preferably alanine, phenylalanine, valine, leucine, or serine;

when L is a di-hexapeptide formed by condensing amino acids, the amino acids are glycine, alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine or histidine, preferably glycine, alanine, valine, serine or phenylalanine;

when X and Y are independently selected from the group consisting of a di-hexapeptide formed by the condensation of amino acids, said di-hexapeptide is glycine-, glycine-alanine, alanine-alanine, glycine-valine, glycine-serine, or glycine-phenylalanine;

when X and Y are independently selected from amino acids, said amino acids are alanine, phenylalanine, methionine, glutamic acid, aspartic acid, leucine and proline, preferably alanine, phenylalanine, valine, leucine or serine;

when X and Y are independently selected from the group consisting of di-hexapeptides formed by the condensation of amino acids that are glycine, alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, or histidine, preferably glycine, alanine, valine, serine, or phenylalanine;

when X and Y are independently selected from the group consisting of a di-hexapeptide formed by the condensation of amino acids, said di-hexapeptide is glycine-, glycine-alanine, alanine-alanine, glycine-valine, glycine-serine, or glycine-phenylalanine;

when R is¹Is unsubstituted or R^1-1Substituted C_1-6When it is alkyl, said R^1-1The number of (a) is 1,2 or 3;

when R is¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl or isopropyl, preferably methyl, ethyl or propyl;

when R is¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl or isopropyl, preferably methyl;

when R is¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isobutyl or isopropyl;

when R is¹Is unsubstituted or R^1-1Substituted C_1-6When it is alkyl, said R^1-1Substituted C_1-6Alkyl is

When R is¹Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

when R is¹Is C_6-12Aryl radical- (C)_1-4Alkyl) -said C_6-12Aryl radical- (C)_1-4Alkyl) -is phenyl- (C)_1-2Alkyl groups);

when R is¹Is C_6-12Aryl radical- (C)_1-4Alkyl) -said C_6-12Aryl radical- (C)_1-4Alkyl) -is benzyl or phenethyl;

n is an integer of 0 to 200;

when R is²Is C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isopropyl or isobutyl, preferably methyl or ethyl;

when R is³Is C_1-6When alkyl, said C_1-6The alkyl is methyl, ethyl, propyl, butyl, sec-butyl or iso-butylPropyl or isobutyl, preferably methyl or ethyl.

In one embodiment, certain groups of the compounds of formula I and II are as defined below (undefined groups are as in any of the preceding embodiments):

l is

Amino acids, or, di-hexapeptides formed by amino acid condensation;

x is

Amino acids or amino acids condensed to form di-hexapeptides;

y is

Amino acids or amino acids condensed to form di-hexapeptides;

t is 1 or 2;

R¹is hydrogen, unsubstituted or R^1-1Substituted C_1-6Alkyl radical, C_6-10Aryl or C_3-10A cycloalkyl group;

R^1-1is a hydroxyl group;

n is an integer of 0 to 200;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-a linear polyethylene glycol;

r is-O (R)²)；

R²Is hydrogen or C_1-6An alkyl group.

In one embodiment, certain groups of the compounds of formula I and II are as defined below (undefined groups are as in any of the preceding embodiments):

l is

Amino acids or amino acids condensed to form di-hexapeptides;

x is

Amino acids or amino acids condensed to form di-hexapeptides;

y is

Amino acids or amino acids condensed to form di-hexapeptides;

t is 1 or 2;

R¹is hydrogen, unsubstituted or R^1-1Substituted C_1-6Alkyl or C_3-10A cycloalkyl group;

R^1-1is a hydroxyl group;

n is an integer of 0 to 100;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-a linear polyethylene glycol;

r is-O (R)²)；

R²Is hydrogen or C_1-6An alkyl group.

In one embodiment, certain groups of the compounds of formula I and II are as defined below (undefined groups are as in any of the preceding embodiments):

in one embodiment, the compounds of formula i and ii may be of any of the following structures:

the invention also provides a preparation method of the compound shown in the formula I, and the synthesis comprises the following steps: in a solvent, under the action of alkali and a condensing agent, carrying out condensation reaction on a compound shown as a formula III and a compound shown as a formula IV as shown in the specification;

wherein L and R are as defined above.

The invention also provides a preparation method of the compound shown in the formula I, and a preparation method of the compound shown in the formula II, wherein the synthesis comprises the following steps: in a solvent, under the action of alkali and a condensing agent, carrying out a condensation reaction as shown in the specification on a compound shown in a formula V and a compound shown in a formula III;

wherein X and Y are as defined above.

The invention also provides application of the compound shown as the formula I, pharmaceutically acceptable salts, metabolic precursors, metabolic products or isomers thereof in preparing medicaments, and the medicaments can be used for preventing and/or treating diabetic complications and pulmonary fibrosis.

The invention also provides a pharmaceutical composition comprising a compound of formula i and formula ii, pharmaceutically acceptable salts thereof, metabolic precursors thereof, metabolites thereof or isomers thereof, as claimed in any one of claims 1 to 7, and a pharmaceutically acceptable adjuvant.

In the pharmaceutical composition, the compound represented by formula i, a pharmaceutically acceptable salt thereof, a metabolic precursor thereof, a metabolic product thereof, or an isomer thereof may be used in a therapeutically effective amount.

The invention also provides application of the pharmaceutical composition in preparation of medicaments, and the medicaments can be used for preventing and/or treating diabetic complications and pulmonary fibrosis.

The pharmaceutical excipients can be those widely used in the field of pharmaceutical production. The adjuvants are used to provide a safe, stable and functional pharmaceutical composition, and may also provide methods for dissolving the active ingredient at a desired rate or for promoting effective absorption of the active ingredient after administration of the composition by a subject. The pharmaceutical excipients may be inert fillers or provide certain functions, such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients may include one or more of the following excipients: binders, suspending agents, emulsifying agents, diluents, fillers, granulating agents, adhesives, disintegrating agents, lubricants, anti-adherents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, reinforcing agents, adsorbents, buffering agents, chelating agents, preservatives, coloring agents, flavoring agents and sweetening agents.

The pharmaceutical compositions of the present invention may be prepared according to the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, levigating, encapsulating, entrapping or lyophilizing processes.

The pharmaceutical compositions of the present invention may be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implant, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical compositions of the present invention may also be in a controlled release or delayed release dosage form (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs and solutions. Examples of topical formulations include, but are not limited to, creams, gels, ointments, creams, patches, pastes, foams, lotions, drops, or serum preparations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry preparations which can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic formulations; aerosol: such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from the compounds of the present invention found to have particular substituents, with relatively nontoxic acids or bases. When compounds of the invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts with inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, bisulfate, hydroiodic acid, phosphorous acid, and the like; and salts of organic acids including acids such as acetic, propionic, isobutyric, maleic, malonic, benzoic, succinic, suberic, trans-succinic, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, and methanesulfonic acids and the like; also included are Salts of amino acids (e.g., arginine, etc.), and Salts of organic acids such as glucuronic acid (see Berge et al, "Pharmaceutical Salts," Journal of Pharmaceutical Science 66:1-19 (1977)). Certain specific compounds of the invention contain both basic and acidic functionalities and can thus be converted to any base or acid addition salt. Preferably, the neutral form of the compound is regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner. The parent form of the compound differs from its various salt forms in certain physical properties, such as solubility in polar solvents.

The "pharmaceutically acceptable salts" of the present invention can be synthesized from the parent compound containing an acid or base by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

The term "metabolite" refers to a pharmaceutically active product of a compound of formula I or a salt thereof produced by in vivo metabolism. Such products may result from, for example, oxidation, reduction, hydrolysis, amidation, deamidation, esterification, deesterification, glucuronidation (glucuronidation), enzymatic cleavage, etc. of the administered compound. Accordingly, the invention includes metabolites of the compounds of the invention, including compounds produced by a method comprising contacting a compound of the invention with a mammal for a period of time sufficient to obtain a metabolite thereof.

Identification of metabolites is typically by making a radiolabel of a compound of the invention (e.g., 14C or³H) Isotopes, which are administered parenterally to an animal, such as a rat, mouse, guinea pig, monkey, or human at detectable doses (e.g., greater than about 0.5mg/kg), allowing sufficient time for metabolism to occur (typically about 30 seconds to 30 hours) and for their conversion products to be isolated from urine, blood or other biological samples. These products are easy to isolate because they are labelled (others are isolated by using antibodies capable of binding to epitopes present in the metabolite). Metabolite structure is determined in a conventional manner, e.g., by MS, LC/MS or NMR analysis. Typically, the analysis of metabolites is in routine drug metabolism studies well known to those skilled in the artThe method is carried out in the same way. So long as the metabolite products are not otherwise detectable in vivo, they can be used in assays for therapeutic dosing of the compounds of the invention. The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compound may be labelled with a radioisotope, such as tritium (A), (B), (C) and C)³H) Iodine-125 (¹²⁵I) Or C-14(¹⁴C) In that respect All permutations of the isotopic composition of the compounds of the present invention, radioactive or otherwise, are intended to be encompassed within the scope of the present invention.

The term "isomer" refers to two or more compounds that are identical in molecular composition but differ in structure and properties.

It will be understood by those skilled in the art that the term "" as used in the structural formulae describing groups herein means that the corresponding group is attached via this site to other fragments, groups in the compounds of formula I and II, according to convention used in the art.

The "substitution" in the present invention may be one or more, and when there are a plurality of "substitutions", the "substitutions" may be the same or different.

The term "plurality" means 2,3, 4 or 5.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "alkyl" refers to a straight or branched chain alkyl group having the indicated number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like.

The term "alkoxy" refers to the group-O-R^YWherein R is^YIs an alkyl group as defined above.

The term "cycloalkyl" refers to a saturated monocyclic or polycyclic alkyl group. The monocyclic cycloalkyl group is preferably a monovalent saturated cyclic alkyl group having 3 to 7 ring carbon atoms, more preferably 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. Each ring of the polycyclic cycloalkyl is saturated and can be a bicyclic or tricyclic cycloalkyl having 4 to 10 carbon atoms.

The term "heterocycloalkyl" refers to a saturated monocyclic or polycyclic group having a heteroatom. The monocyclic ring preferably contains 1,2 or 3-7 membered saturated monocyclic heterocycloalkyl independently selected from N, O and S, examples of which include but are not limited to: pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrrolyl, azetidinyl, thiazolidinyl, oxazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, azepanyl, diazepanyl, oxazepanyl, dioxolanyl, dioxanyl, and the like. The polycyclic rings are preferably 8-10 membered saturated polycyclic heterocycloalkyl containing 1,2 or 3 rings independently selected from N, O and S, which may be bicyclic or tricyclic, examples include, but are not limited to, octahydropyrrolo [1,2-a ] pyrazinyl, (1R, 5S) -3, 8-diazabicyclo [3.2.1] octyl.

The term "aryl" refers to an aromatic group having the indicated number of carbon atoms, preferably a monocyclic, bicyclic or tricyclic aromatic group, each of which, when bicyclic or tricyclic, satisfies the huckel rule. C of the invention_6-10The aryl group of (b) means an aromatic group having 6 to 10 carbon atoms, such as phenyl or naphthyl.

The term "heteroaryl" refers to an aromatic group containing a heteroatom, preferably an aromatic 5-6 membered monocyclic or 9-10 membered bicyclic ring containing 1,2 or 3 members independently selected from nitrogen, oxygen and sulfur. The 5-to 6-membered monocyclic ring includes, but is not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, 1,2, 3-triazolyl, 1,2, 4-triazolyl, furazanyl, 1,2, 3-oxadiazolyl, 1,2, 4-oxadiazole, 1,2, 5-oxadiazole, 1,3, 4-oxadiazole, thiadiazolyl, dithiazolyl, tetrazolyl, pyridyl, pyranyl, thiapyranyl, diazinyl, pyrazinyl, pyrimidinyl, pyridazinyl, oxazinyl, thiazinyl, dioxinyl, dithiinyl, 1,2, 3-triazinyl, 1,2, 4-triazinyl, 1,3, 5-triazinyl, or tetrazinyl. The 9-to 10-membered bicyclic ring includes but is not limited to benzimidazolyl, indolyl, indazolyl, benzofuranyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, quinolyl and isoquinolyl.

The above preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention without departing from the common general knowledge in the art.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows:

(1) the medicine water solubility of the PEGs-modified vincamine is enhanced;

(2) the PEGylated vincamine prodrug has good therapeutic effect on diabetic complications including diabetic peripheral neuropathy and diabetic foot;

(3) the PEGylated vincamine prodrug has good therapeutic effect on pulmonary fibrosis.

Drawings

FIG. 1 is an immunofluorescence and quantitative bar chart of vincamine derivative Vin3 capable of promoting the growth of peripheral sensory neuron axons of type 1 diabetic mice.

FIG. 2 is an immunofluorescence and quantitative bar chart of vincamine derivative Vin3 capable of significantly promoting the growth of peripheral sensory neuron axons of type 2 diabetic mice.

Fig. 3 is a photograph showing that the vincamine derivative Vin3 can obviously promote the healing of foot ulcer wounds of diabetic foot rats.

Fig. 4 is a CT picture that the vincamine derivative Vin3 can obviously improve the degree of pulmonary fibrosis of mice.

Detailed Description

The present invention is further illustrated below with reference to specific examples, which should not be construed as limiting the invention.

Example 1: synthesis of Compound S1

The method comprises the following steps: synthesis of vincamine demethylation product

1.0g (2.8mmol) of vincamine was dissolved in 10mL of NaOH solution (1mol/L) at room temperature, stirred, the solution gradually changed from turbid to clear, and the progress of the reaction was checked by TLC spot plate. After the reaction of the vincamine is finished, the reaction solution is placed at 0 ℃ and the pH value is adjusted to 3 by using dilute hydrochloric acid, then methanol is removed by rotary evaporation, filtration is carried out, a filter cake is collected and dried in vacuum, and the crude product of the vincamine demethylate is obtained and is directly put into the next step.

¹H NMR(300MHz,Chloroform-d)δ11.69(s,1H),7.53–7.46(m,1H),7.17–7.06(m, 3H),4.64(s,1H),3.92(s,1H),3.39–3.21(m,2H),3.07–2.92(m,1H),2.65–2.45(m,3H), 2.33–2.09(m,3H),1.78–1.62(m,2H),1.53–1.34(m,3H),0.91(t,J＝7.6Hz,3H)。

Step two: synthesis of vincamine glycinate

200mg (0.59mmol) of vincamine demethylation product, 99mg (0.59mmol) of glycine tert-butyl ester hydrochloride, 192mg (1.00mmol) of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI), and 103mg (0.76mmol) of 1-hydroxybenzotriazole (HOBt) were dissolved in 8mL of anhydrous N, N-Dimethylformamide (DMF) at room temperature, stirred, added with 0.4mL (2.94mmol) of triethylamine, and reacted under nitrogen at room temperature for 10 hours. DMF was removed by rotary evaporation, dissolved in dichloromethane, extracted (3X 20mL), washed with saturated brine (1X 20mL), and dried over anhydrous magnesium sulfate. The solution is separated by spin-drying silica gel column chromatography (dichloromethane: methanol 60:1) to obtain the vincamine glycine protection product of 160 mg. The vincamine glycine protected product, 160mg (0.35mmol), was dissolved in 10mL trifluoroacetic acid in dichloromethane (50% v/v) at 0 deg.C and stirred for 1 hour, then warmed to room temperature and stirred at room temperature for 3 hours. And (3) performing rotary evaporation to remove the dichloromethane and the trifluoroacetic acid, adding 20mL of diethyl ether, stirring for 1 hour, filtering, collecting a filter cake, and performing vacuum drying to obtain 110mg of a vincamine glycine crude product, and directly putting the vincamine glycine crude product into the next step.

¹H NMR(300MHz,Methanol-d4)δ7.58–7.45(m,2H),7.31–7.17(m,2H),5.79(s, 1H),5.06(s,1H),4.15(q,J＝17.5Hz,2H),3.73(td,J＝12.4,5.5Hz,1H),3.64–3.56(m,2H), 3.26–2.91(m,3H),2.37(dd,J＝17.0,5.0Hz,1H),2.07–1.66(m,5H),1.29–1.14(m,2H), 1.09(t,J＝7.4Hz,3H)。

Step three: synthesis of Compound S1

200mg (0.50mmol) of vincamine glycinate, 61mg (0.50mmol) of diethylene glycol monomethyl ether, 156mg (0.76mmol) of Dicyclohexylcarbodiimide (DCC) and 11mg (0.05mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) are dissolved in 2mL of dichloromethane at room temperature and reacted for 12 hours at normal temperature under the protection of nitrogen. Insoluble matters are removed by suction filtration, water is added into the filtrate, dichloromethane is used for extraction (5mL multiplied by 3), organic phases are combined, saturated saline (2mL) is washed for 1 time, anhydrous sodium sulfate is dried, filtration is carried out, the solvent is evaporated to dryness, sand is prepared, and column chromatography separation and purification (DCM: MeOH ═ 60: 1-40: 1) are carried out, so as to obtain white solid S1187 mg.

¹H NMR(300MHz,Chloroform-d)δ7.45(dd,J＝6.5,2.1Hz,1H),7.30(dd,J＝6.9,1.8 Hz,1H),7.20–7.07(m,2H),5.73(s,1H),4.41–4.14(m,5H),3.82(s,1H),3.77–3.70(t,J＝ 4.7Hz,2H),3.67–3.61(m,2H),3.58–3.51(m,2H),3.37(s,3H),3.36–3.19(m,2H),3.09– 2.95(m,1H),2.68–2.60(m,2H),2.51(dd,J＝16.3,2.8Hz,1H),2.00–1.80(m,3H),1.80– 1.62(m,1H),1.56–1.38(m,2H),1.38–1.20(m,2H),1.00(t,J＝7.5Hz,3H)。

The compounds S2 to S4 in the following examples 2 to 4 can be obtained by the synthesis method of example 1, and only the corresponding raw materials need to be replaced.

Example 2: synthesis of Compound S2

¹H NMR(300MHz,Chloroform-d)δ7.46(dd,J＝6.4,2.1Hz,1H),7.30(dd,J＝6.9,1.8 Hz,1H),7.22–7.07(m,2H),5.75(s,1H),4.45–4.17(m,5H),3.83(s,1H),3.78–3.70(t,J＝4.6Hz,2H),3.69–3.62(m,6H),3.59–3.48(m,2H),3.37(s,3H),3.36–3.20(m,2H),3.11– 2.96(m,1H),2.68–2.61(m,2H),2.52(dd,J＝16.2,2.8Hz,1H),2.03–1.79(m,3H),1.79– 1.61(m,1H),1.58–1.39(m,2H),1.39–1.21(m,2H),1.01(t,J＝7.4Hz,3H)。

Example 3: synthesis of Compound S3

¹H NMR(300MHz,Chloroform-d)δ7.45(dd,J＝6.5,2.1Hz,1H),7.30(dd,J＝6.8,1.8 Hz,1H),7.20–7.07(m,2H),5.75(s,1H),4.42–4.14(m,5H),3.82(s,1H),3.78–3.69(t,J＝ 4.7Hz,2H),3.67–3.58(m,10H),3.56–3.45(m,2H),3.38(s,3H),3.37–3.20(m,2H),3.10– 2.95(m,1H),2.69–2.61(m,2H),2.51(dd,J＝16.2,2.8Hz,1H),2.02–1.81(m,3H),1.81– 1.62(m,1H),1.60–1.40(m,2H),1.40–1.19(m,2H),1.01(t,J＝7.5Hz,3H)。

Example 4: synthesis of Compound S4

¹H NMR(300MHz,Chloroform-d)δ7.45(dd,J＝6.5,2.1Hz,1H),7.31(dd,J＝6.9,1.8 Hz,1H),7.20–7.05(m,2H),5.74(s,1H),4.40–4.15(m,5H),3.82(s,1H),3.76–3.70(t,J＝ 4.6Hz,2H),3.68–3.58(m,14H),3.57–3.42(m,2H),3.37(s,3H),3.36–3.20(m,2H),3.11– 2.96(m,1H),2.67–2.60(m,2H),2.51(dd,J＝16.3,2.8Hz,1H),2.00–1.80(m,3H),1.79– 1.62(m,1H),1.56–1.40(m,2H),1.39–1.21(m,2H),1.00(t,J＝7.5Hz,3H)。

Example 5: synthesis of Compound S5

200mg (0.50mmol, synthesis method shown in example 1) of vincamine glycinate, 1008mg (0.50mmol) of monomethoxyhydroxypolyethylene glycol (MPEG-OH, average molecular weight 2000), 156mg (0.76mmol) of Dicyclohexylcarbodiimide (DCC), 11mg (0.05mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) are dissolved in 10mL of dichloromethane at room temperature, and reacted for 12 hours at normal temperature under the protection of nitrogen. And (3) carrying out suction filtration to remove insoluble substances, carrying out rotary evaporation on the filtrate to remove dichloromethane, adding 250mL of diethyl ether, stirring for 1 hour, filtering, collecting a filter cake, and carrying out vacuum drying to obtain a crude product. The crude product was dialyzed to give S5610 mg as a white solid.

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.13(m,2H),5.75(s,1H),4.42–4.13(m,4H),3.90–3.81(m,3H),3.78–3.50(m), 3.43–3.34(m,6H),3.14–2.86(m,3H),2.41–1.77(m,7H),1.70–1.53(m,2H),1.30–1.09 (m,2H),1.04(t,J＝7.3Hz,3H)。

The compounds S6 to S22, S27 to S30 and S39 to S42 in examples 6 to 22, 27 to 30 and 39 to 42 were all obtained by the synthesis method of example 5, and they were replaced with the corresponding amino acid (hydroxypropionic acid) and monomethoxyhydroxypolyethylene glycol of different molecular weights.

Example 6: synthesis of Compound S6

¹H NMR(300MHz,Chloroform-d)δ7.47(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.12(m,2H),5.76(s,1H),4.41–4.13(m,4H),3.90–3.80(m,3H),3.80–3.49(m), 3.43–3.34(m,6H),3.14–2.85(m,3H),2.41–1.78(m,7H),1.70–1.53(m,2H),1.30–1.10 (m,2H),1.04(t,J＝7.3Hz,3H)。

Example 7: synthesis of Compound S7

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.2Hz,1H),7.39(d,J＝8.4Hz,1H), 7.24–7.13(m,2H),5.76(s,1H),4.42–4.12(m,4H),3.91–3.81(m,3H),3.77–3.49(m), 3.43–3.33(m,6H),3.14–2.86(m,3H),2.42–1.77(m,7H),1.70–1.53(m,2H),1.30–1.09 (m,2H),1.04(t,J＝7.3Hz,3H)。

Example 8: synthesis of Compound S8

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.75(s,1H),4.42–4.13(m,4H),3.91–3.81(m,3H),3.80–3.49(m), 3.43–3.33(m,6H),3.15–2.86(m,3H),2.41–1.76(m,7H),1.70–1.53(m,2H),1.30–1.09 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 9: synthesis of Compound S9

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.14(m,2H),5.76(s,1H),4.42–4.12(m,4H),3.90–3.82(m,3H),3.80–3.50(m), 3.43–3.34(m,6H),3.14–2.86(m,3H),2.41–1.76(m,7H),1.71–1.53(m,2H),1.30–1.10 (m,2H),1.04(t,J＝7.3Hz,3H)。

Example 10: synthesis of Compound S10

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.2Hz,1H),7.38(d,J＝8.4Hz,1H), 7.25–7.12(m,2H),5.75(s,1H),4.43–4.13(m,4H),3.90–3.80(m,3H),3.81–3.49(m), 3.44–3.34(m,6H),3.14–2.85(m,3H),2.41–1.77(m,7H),1.70–1.54(m,2H),1.30–1.10 (m,2H),1.04(t,J＝7.3Hz,3H)。

Example 11: synthesis of Compound S11

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.75(s,1H),4.41–4.13(m,4H),3.91–3.81(m,3H),3.82–3.49(m), 3.43–3.33(m,6H),3.14–2.86(m,3H),2.40–1.77(m,7H),1.71–1.53(m,2H),1.30–1.09 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 12: synthesis of Compound S12

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.40(d,J＝8.3Hz,1H), 7.27–7.13(m,2H),5.76(s,1H),4.42–4.13(m,3H),3.90–3.82(m,3H),3.79–3.50(m), 3.43–3.33(m,6H),3.14–2.85(m,3H),2.41–1.76(m,7H),1.71–1.53(m,2H),1.37(d,J＝ 6.8Hz,3H),1.30–1.10(m,2H),1.03(t,J＝7.3Hz,3H)。

Example 13: synthesis of Compound S13

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.4Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.76(s,1H),4.43–4.13(m,3H),3.90–3.83(m,3H),3.79–3.49(m), 3.43–3.32(m,6H),3.13–2.85(m,3H),2.41–1.75(m,8H),1.71–1.53(m,2H),1.30–1.09 (m,2H),1.03(t,J＝7.3Hz,3H),0.96(dd,J＝6.6,24.9Hz,6H)。

Example 14: synthesis of Compound S14

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.4Hz,1H),7.40(d,J＝8.3Hz,1H), 7.28–7.13(m,2H),5.76(s,1H),4.42–4.14(m,3H),3.91–3.83(m,3H),3.79–3.51(m), 3.44–3.33(m,6H),3.14–2.86(m,5H),2.42–1.76(m,7H),1.71–1.54(m,2H),1.31–1.10 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 15: synthesis of Compound S15

¹H NMR(300MHz,Chloroform-d)δ7.42–7.34(m,2H),7.30–7.19(m,5H),7.16– 7.10(m,2H),5.75(s,1H),4.43–4.14(m,3H),3.90–3.83(m,3H),3.79–3.52(m),3.43– 3.33(m,6H),3.14–2.85(m,5H),2.41–1.76(m,7H),1.71–1.53(m,2H),1.30–1.10(m, 2H),1.04(t,J＝7.3Hz,3H)。

Example 16: synthesis of Compound S16

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.12(m,2H),5.76(s,1H),4.44–4.12(m,3H),3.91–3.83(m,3H),3.79–3.48(m), 3.43–3.33(m,6H),3.14–2.85(m,3H),2.41–1.74(m,8H),1.71–1.52(m,4H),1.30–1.10 (m,2H),1.03(t,J＝7.3Hz,3H),0.96–0.83(m,6H)。

Example 17: synthesis of Compound S17

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.13(m,2H),5.75(s,1H),4.43–4.13(m,4H),3.90–3.81(m,3H),3.79–3.50(m), 3.43–3.33(m,6H),3.15–2.86(m,3H),2.40–1.77(m,7H),1.70–1.52(m,2H),1.32–1.09 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 18: synthesis of Compound S18

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.39(d,J＝8.3Hz,1H), 7.27–7.14(m,2H),5.76(s,1H),4.43–4.13(m,3H),3.91–3.82(m,3H),3.79–3.49(m), 3.43–3.33(m,6H),3.14–2.86(m,3H),2.41–1.77(m,7H),1.71–1.53(m,2H),1.36(d,J＝ 6.8Hz,3H),1.30–1.09(m,2H),1.03(t,J＝7.3Hz,3H)。

Example 19: synthesis of Compound S19

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.39(d,J＝8.3Hz,1H), 7.27–7.14(m,2H),5.76(s,1H),4.43–4.13(m,3H),3.91–3.82(m,3H),3.79–3.49(m), 3.43–3.33(m,6H),3.15–2.85(m,3H),2.42–1.76(m,8H),1.71–1.54(m,2H),1.31–1.10 (m,2H),1.03(t,J＝7.3Hz,3H),0.96(dd,J＝6.6,24.9Hz,6H)。

Example 20: synthesis of Compound S20

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.40(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.75(s,1H),4.41–4.15(m,3H),3.90–3.83(m,3H),3.79–3.50(m), 3.44–3.34(m,6H),3.14–2.86(m,5H),2.43–1.76(m,7H),1.71–1.53(m,2H),1.31–1.10 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 21: synthesis of Compound S21

¹H NMR(300MHz,Chloroform-d)δ7.43–7.34(m,2H),7.31–7.19(m,5H),7.16– 7.10(m,2H),5.75(s,1H),4.44–4.14(m,3H),3.90–3.82(m,3H),3.79–3.51(m),3.43– 3.32(m,6H),3.15–2.85(m,5H),2.41–1.75(m,7H),1.72–1.53(m,2H),1.31–1.10(m, 2H),1.04(t,J＝7.3Hz,3H)。

Example 22: synthesis of Compound S22

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.39(d,J＝8.3Hz,1H), 7.24–7.12(m,2H),5.76(s,1H),4.43–4.12(m,3H),3.90–3.83(m,3H),3.79–3.47(m), 3.44–3.33(m,6H),3.14–2.86(m,3H),2.41–1.73(m,8H),1.71–1.51(m,4H),1.30–1.09 (m,2H),1.03(t,J＝7.3Hz,3H),0.95–0.83(m,6H)。

Example 23: synthesis of Compound S23

200mg (0.50mmol, see example 1 for synthesis method), 1008mg (0.50mmol) of dihydroxypolyethylene glycol (OH-PEG-OH, average molecular weight 2000), 156mg (0.76mmol) of Dicyclohexylcarbodiimide (DCC), 11mg (0.05mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) are dissolved in 10mL of dichloromethane at room temperature, and reacted for 12 hours at normal temperature under the protection of nitrogen. And (3) carrying out suction filtration to remove insoluble substances, carrying out rotary evaporation on the filtrate to remove dichloromethane, adding 250mL of diethyl ether, stirring for 1 hour, filtering, collecting a filter cake, and carrying out vacuum drying to obtain a crude product. The crude product was dialyzed to give S23600 mg as a white solid.

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.76(s,1H),4.41–4.13(m,4H),3.91–3.81(m,3H),3.78–3.49(m), 3.43–3.33(m,6H),3.15–2.86(m,3H),2.42–1.77(m,7H),1.70–1.53(m,2H),1.30–1.08 (m,2H),1.04(t,J＝7.3Hz,3H)。

The compounds S24 to S26 in the following examples 24 to 26 can be obtained by the synthesis method of example 23 by replacing with corresponding amino acids and dihydroxy polyethylene glycols of different molecular weights.

Example 24: synthesis of Compound S24

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.75(s,1H),4.42–4.14(m,3H),3.91–3.80(m,3H),3.78–3.49(m), 3.43–3.34(m,6H),3.14–2.85(m,3H),2.41–1.76(m,7H),1.70–1.53(m,2H),1.37(d,J＝ 6.8Hz,3H),1.30–1.10(m,2H),1.04(t,J＝7.3Hz,3H)。

Example 25: synthesis of Compound S25

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.75(s,1H),4.43–4.13(m,4H),3.91–3.81(m,3H),3.78–3.50(m), 3.43–3.33(m,6H),3.14–2.85(m,3H),2.42–1.77(m,7H),1.70–1.52(m,2H),1.31–1.09 (m,2H),1.04(t,J＝7.3Hz,3H)。

Example 26: synthesis of Compound S26

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.75(s,1H),4.43–4.13(m,3H),3.91–3.81(m,3H),3.78–3.49(m), 3.43–3.33(m,6H),3.14–2.86(m,3H),2.42–1.77(m,7H),1.70–1.52(m,2H),1.37(d,J＝ 6.8Hz,3H),1.30–1.10(m,2H),1.04(t,J＝7.3Hz,3H)。

Example 27: synthesis of Compound S27

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.40(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.76(s,1H),4.43–4.12(m,4H),3.91–3.81(m,3H),3.78–3.49(m), 3.43–3.33(m,6H),3.21(s,3H),3.15–2.86(m,3H),2.42–1.77(m,7H),1.71–1.53(m,2H), 1.31–1.09(m,2H),1.04(t,J＝7.3Hz,3H)。

Example 28: synthesis of Compound S28

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.40(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.76(s,1H),4.43–4.12(m,4H),3.91–3.81(m,3H),3.78–3.49(m), 3.43–3.32(m,8H),3.15–2.86(m,3H),2.42–1.77(m,7H),1.71–1.53(m,4H),1.31–1.09 (m,2H),1.04(t,J＝7.3Hz,3H),0.88(t,J＝7.7Hz,3H)。

Example 29: synthesis of Compound S29

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.13(m,2H),5.76(s,1H),4.43–4.11(m,4H),3.90–3.82(m,3H),3.79–3.50(m), 3.43–3.31(m,6H),3.26–3.24(m,1H),3.15–2.86(m,3H),2.43–1.76(m,7H),1.71–1.53 (m,2H),1.34–1.09(m,6H),1.04(t,J＝7.3Hz,3H)。

Example 30: synthesis of Compound S30

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.40(d,J＝8.3Hz,1H), 7.36–7.28(m,2H),7.26–7.13(m,5H),5.76(s,1H),4.42–4.12(m,4H),3.91–3.80(m,3H), 3.78–3.50(m),3.44–3.33(m,6H),3.14–2.85(m,3H),2.42–1.76(m,7H),1.72–1.53(m, 2H),1.31–1.10(m,2H),1.04(t,J＝7.3Hz,3H)。

Example 31: synthesis of Compound S31

Fmoc-protected glycine (845mg, 2.85mmol) was dissolved in anhydrous CH₂Cl₂(30mL), L-valine tert-butyl ester hydrochloride (659mg, 1.42mmol), HATU (2.16g, 5.70mmol), HOAt (1.38g, 10.15mmol), DIPEA (1.98mL, 11.4mmol) were added in sequence at room temperature, the reaction was allowed to warm to room temperature for 7 hours, the progress of the reaction was checked by TLC plates, and after completion of the reaction, saturated NH was added₄Cl solution is extracted and killed, and the organic phase is sequentially added with saturated NaHCO₃Washing with water and saturated salt water, and washing with anhydrous Na₂SO₄Drying, filtering, evaporating the solvent under reduced pressure, and then carrying out column chromatography to obtain white Fmoc-protected dipeptide solid. The resulting Fmoc-protected dipeptide (1.11g, 2.45mmol) was dissolved in MeCN (200mL), piperidine (10mL) was added at room temperature for reaction for 3 hours, the solvent was distilled off under reduced pressure, and the resulting Fmoc-removed dipeptide was purified by column chromatography to give a white solid compound (270mg, 41%) of Fmoc-removed dipeptide which was reacted with the vincamine demethylate product and monomethoxyhydroxypolyethylene glycol, respectively, according to the methods of step two and step three in example 1, to give compound S31.

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.26–7.13(m,2H),5.75(s,1H),4.43–4.12(m,5H),3.91–3.81(m,3H),3.78–3.50(m), 3.43–3.33(m,6H),3.15–2.86(m,3H),2.41–1.76(m,8H),1.70–1.52(m,2H),1.30–1.09 (m,2H),1.06(dd,J＝6.7,25.0Hz,6H),1.03(t,J＝7.3Hz,3H)。

The compounds S32-S38 in the following examples 32-38 can be obtained by the synthesis method of example 31 by replacing corresponding amino acids and monomethoxyhydroxypolyethylene glycol with different molecular weights.

Example 32: synthesis of Compound S32

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.38(d,J＝8.3Hz,1H), 7.30–7.13(m,7H),5.74(s,1H),4.80(m,1H),4.41–4.13(m,4H),3.91–3.81(m,3H),3.79 –3.50(m),3.43–3.33(m,6H),3.15–2.86(m,5H),2.42–1.77(m,7H),1.71–1.53(m,2H), 1.30–1.10(m,2H),1.04(t,J＝7.3Hz,3H)。

Example 33: synthesis of Compound S33

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.40(d,J＝8.3Hz,1H), 7.27–7.12(m,2H),5.76(s,1H),4.43–4.13(m,4H),3.91–3.82(m,3H),3.79–3.51(m), 3.43–3.33(m,6H),3.15–2.85(m,3H),2.41–1.77(m,7H),1.71–1.53(m,2H),1.37(d,J＝ 6.8Hz,3H),1.35(d,J＝6.8Hz,3H),1.30–1.10(m,2H),1.03(t,J＝7.3Hz,3H)。

Example 34: synthesis of Compound S34

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.13(m,2H),5.75(s,1H),4.46–4.13(m,5H),3.91–3.80(m,5H),3.78–3.50(m), 3.44–3.34(m,6H),3.14–2.86(m,3H),2.41–1.76(m,7H),1.70–1.54(m,2H),1.30–1.09 (m,2H),1.04(t,J＝7.3Hz,3H)。

Example 35: synthesis of Compound S35

¹H NMR(300MHz,Chloroform-d)δ7.46(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.13(m,2H),5.75(s,1H),4.42–4.13(m,5H),3.90–3.81(m,3H),3.78–3.49(m), 3.43–3.32(m,6H),3.15–2.86(m,3H),2.42–1.76(m,8H),1.70–1.52(m,2H),1.30–1.09 (m,2H),1.06(dd,J＝6.7,25.0Hz,6H),1.03(t,J＝7.3Hz,3H)。

Example 36: synthesis of Compound S36

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.38(d,J＝8.3Hz,1H), 7.30–7.14(m,7H),5.74(s,1H),4.80(m,1H),4.40–4.13(m,4H),3.92–3.81(m,3H),3.79 –3.50(m),3.43–3.34(m,6H),3.16–2.86(m,5H),2.43–1.77(m,7H),1.71–1.53(m,2H), 1.30–1.10(m,2H),1.04(t,J＝7.3Hz,3H)。

Example 37: synthesis of Compound S37

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.4Hz,1H),7.40(d,J＝8.3Hz,1H), 7.27–7.12(m,2H),5.76(s,1H),4.44–4.13(m,4H),3.91–3.82(m,3H),3.79–3.50(m), 3.43–3.33(m,6H),3.15–2.85(m,3H),2.41–1.76(m,7H),1.71–1.53(m,2H),1.37(d,J＝ 6.8Hz,3H),1.35(d,J＝6.8Hz,3H),1.31–1.10(m,2H),1.03(t,J＝7.3Hz,3H)。

Example 38: synthesis of Compound S38

¹H NMR(300MHz,Chloroform-d)δ7.45(d,J＝7.2Hz,1H),7.39(d,J＝8.3Hz,1H), 7.25–7.13(m,2H),5.75(s,1H),4.44–4.13(m,5H),3.91–3.80(m,5H),3.78–3.49(m), 3.44–3.34(m,6H),3.14–2.86(m,3H),2.40–1.76(m,7H),1.70–1.54(m,2H),1.30–1.09 (m,2H),1.04(t,J＝7.3Hz,3H)。

Example 39: synthesis of Compound S39

¹H NMR(300MHz,Chloroform-d)δ7.44(d,J＝7.2Hz,1H),7.38(d,J＝8.3Hz,1H), 7.24–7.13(m,2H),5.75(s,1H),4.55–4.12(m,6H),3.91–3.81(m,3H),3.79–3.50(m), 3.44–3.34(m,6H),3.14–2.85(m,3H),2.40–1.77(m,7H),1.70–1.53(m,2H),1.30–1.10 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 40: synthesis of Compound S40

¹H NMR(300MHz,Chloroform-d)δ7.44(d,J＝7.2Hz,1H),7.38(d,J＝8.3Hz,1H), 7.24–7.13(m,2H),5.75(s,1H),4.54–4.12(m,6H),3.91–3.81(m,3H),3.80–3.50(m), 3.44–3.34(m,6H),3.14–2.85(m,3H),2.40–1.76(m,7H),1.70–1.53(m,2H),1.30–1.09 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 41: synthesis of Compound S41

¹H NMR(300MHz,Chloroform-d)δ7.44(d,J＝7.2Hz,1H),7.38(d,J＝8.3Hz,1H), 7.24–7.13(m,2H),5.75(s,1H),4.55–4.12(m,6H),3.90–3.81(m,3H),3.80–3.50(m), 3.44–3.34(m,6H),3.14–2.85(m,3H),2.40–1.76(m,7H),1.70–1.53(m,2H),1.31–1.10 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 42: synthesis of Compound S42

¹H NMR(300MHz,Chloroform-d)δ7.44(d,J＝7.2Hz,1H),7.38(d,J＝8.3Hz,1H), 7.24–7.13(m,2H),5.76(s,1H),4.54–4.12(m,6H),3.91–3.81(m,3H),3.80–3.49(m), 3.45–3.34(m,6H),3.14–2.85(m,3H),2.40–1.76(m,7H),1.70–1.53(m,2H),1.31–1.10 (m,2H),1.03(t,J＝7.3Hz,3H)。

Example 43: synthesis of Compound S43

440mg (1.10mmol, see example 1 for synthesis method), 1008mg (0.50mmol) of dihydroxypolyethylene glycol (OH-PEG-OH, average molecular weight 2000), 248mg (1.20mmol) of Dicyclohexylcarbodiimide (DCC), 11mg (0.05mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) are dissolved in 10mL of dichloromethane at room temperature, and reacted for 12 hours at normal temperature under the protection of nitrogen. And (3) carrying out suction filtration to remove insoluble substances, carrying out rotary evaporation on the filtrate to remove dichloromethane, adding 250mL of diethyl ether, stirring for 1 hour, filtering, collecting a filter cake, and carrying out vacuum drying to obtain a crude product. The crude product was dialyzed to give S43730 mg as a white solid.

¹H NMR(300MHz,Chloroform-d)δ7.43–7.37(m,2H),7.36–7.28(m,4H),7.22– 7.08(m,2H),5.72(s,2H),4.40–4.14(m,8H),3.87–3.77(m,6H),3.74–3.45(m),3.40– 3.29(m,12H),3.12–2.83(m,6H),2.38–1.73(m,14H),1.68–1.50(m,4H),1.28–1.06(m, 4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

The compounds S44 to S56 in examples 44 to 56 below were all obtained by the synthesis method of example 43, and were replaced with the corresponding amino acids and dihydroxy polyethylene glycols of different molecular weights.

Example 44: synthesis of Compound S44

¹H NMR(300MHz,Chloroform-d)δ7.44–7.37(m,2H),7.36–7.27(m,4H),7.22– 7.09(m,2H),5.72(s,2H),4.41–4.14(m,6H),3.87–3.76(m,6H),3.75–3.45(m),3.40– 3.28(m,12H),3.12–2.82(m,6H),2.37–1.73(m,14H),1.69–1.50(m,4H),1.33(d,J＝6.8 Hz,6H),1.29–1.06(m,4H),0.94(td,J＝1.0,7.2Hz,6H)。

Example 45: synthesis of Compound S45

¹H NMR(300MHz,Chloroform-d)δ7.44–7.37(m,2H),7.37–7.28(m,4H),7.22– 7.08(m,2H),5.73(s,2H),4.41–4.15(m,6H),3.86–3.76(m,6H),3.74–3.45(m),3.39– 3.29(m,12H),3.13–2.83(m,6H),2.38–1.74(m,16H),1.68–1.49(m,4H),1.29–1.07(m, 4H),1.01(m,12H),0.95(t,J＝7.3Hz,3H),0.91(t,J＝7.3Hz,3H)。

Example 46: synthesis of Compound S46

¹H NMR(300MHz,Chloroform-d)δ7.43–7.37(m,2H),7.36–7.28(m,4H),7.23– 7.08(m,2H),5.73(s,2H),4.41–4.15(m,7H),3.86–3.77(m,6H),3.74–3.44(m),3.40– 3.29(m,12H),3.13–2.83(m,6H),2.38–1.73(m,15H),1.68–1.50(m,4H),1.28–1.06(m, 4H),1.02(dd,J＝6.7,25.0Hz,6H),0.94(t,J＝7.3Hz,3H),0.91(t,J＝7.3Hz,3H)。

Example 47: synthesis of Compound S47

¹H NMR(300MHz,Chloroform-d)δ7.44–7.37(m,2H),7.37–7.28(m,4H),7.22– 7.09(m,2H),5.73(s,2H),4.41–4.14(m,8H),3.87–3.76(m,6H),3.75–3.44(m),3.41– 3.29(m,12H),3.12–2.82(m,6H),2.39–1.74(m,14H),1.69–1.50(m,4H),1.28–1.07(m, 4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 48: synthesis of Compound S48

¹H NMR(300MHz,Chloroform-d)δ7.45–7.37(m,2H),7.37–7.27(m,4H),7.22– 7.10(m,2H),5.72(s,2H),4.42–4.14(m,6H),3.86–3.76(m,6H),3.76–3.44(m),3.40– 3.28(m,12H),3.13–2.83(m,6H),2.38–1.74(m,14H),1.70–1.50(m,4H),1.34(d,J＝6.8 Hz,6H),1.29–1.06(m,4H),0.94(td,J＝1.0,7.2Hz,6H)。

Example 49: synthesis of Compound S49

¹H NMR(300MHz,Chloroform-d)δ7.44–7.37(m,2H),7.36–7.29(m,4H),7.22– 7.09(m,2H),5.72(s,2H),4.41–4.14(m,8H),3.88–3.77(m,6H),3.75–3.45(m),3.41–3.29(m,12H),3.12–2.82(m,6H),2.39–1.73(m,14H),1.69–1.50(m,4H),1.28–1.07(m, 4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 50: synthesis of Compound S50

¹H NMR(300MHz,Chloroform-d)δ7.44–7.37(m,2H),7.37–7.28(m,4H),7.23– 7.08(m,2H),5.72(s,2H),4.41–4.14(m,8H),3.88–3.77(m,6H),3.74–3.44(m),3.40– 3.28(m,12H),3.12–2.83(m,6H),2.37–1.73(m,14H),1.69–1.50(m,4H),1.28–1.07(m, 4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 51: synthesis of Compound S51

¹H NMR(300MHz,Chloroform-d)δ7.45–7.37(m,2H),7.37–7.28(m,4H),7.22– 7.10(m,2H),5.72(s,2H),4.42–4.15(m,8H),3.87–3.76(m,6H),3.75–3.44(m),3.40– 3.27(m,12H),3.12–2.82(m,6H),2.38–1.73(m,14H),1.69–1.50(m,4H),1.30–1.06(m, 4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 52: synthesis of Compound S52

¹H NMR(300MHz,Chloroform-d)δ7.45–7.37(m,2H),7.35–7.29(m,4H),7.21– 7.09(m,2H),5.71(s,2H),4.40–4.14(m,7H),4.05–3.93(m,2H),3.88–3.77(m,6H),3.75 –3.45(m),3.41–3.29(m,12H),3.13–2.82(m,6H),2.39–1.74(m,14H),1.69–1.50(m, 4H),1.29–1.07(m,4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 53: synthesis of Compound S53

¹H NMR(300MHz,Chloroform-d)δ7.45–7.37(m,2H),7.36–7.28(m,4H),7.22– 7.09(m,2H),5.72(s,2H),4.41–4.15(m,7H),3.87–3.77(m,6H),3.75–3.45(m),3.41– 3.28(m,12H),3.12–2.82(m,6H),2.39–1.73(m,14H),1.69–1.50(m,4H),1.35(d,J＝6.8 Hz,6H),1.29–1.07(m,4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 54: synthesis of Compound S54

¹H NMR(300MHz,Chloroform-d)δ7.45–7.37(m,2H),7.36–7.29(m,4H),7.22– 7.10(m,2H),5.72(s,2H),4.43–4.14(m,6H),3.89–3.77(m,6H),3.75–3.44(m),3.41– 3.29(m,12H),3.13–2.82(m,6H),2.40–1.73(m,15H),1.69–1.49(m,4H),1.30(d,J＝6.7 Hz,3H),1.28–1.07(m,4H),1.02(dd,J＝6.7,25.0Hz,6H),0.96(t,J＝7.3Hz,3H),0.92(t,J ＝7.3Hz,3H)。

Example 55: synthesis of Compound S55

¹H NMR(300MHz,Chloroform-d)δ7.45–7.37(m,2H),7.35–7.29(m,4H),7.21– 7.10(m,2H),5.71(s,2H),4.40–4.14(m,7H),4.05–3.93(m,2H),3.89–3.77(m,6H),3.75 –3.45(m),3.42–3.29(m,12H),3.13–2.82(m,6H),2.39–1.75(m,14H),1.69–1.50(m, 4H),1.29–1.07(m,4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 56: synthesis of Compound S56

¹H NMR(300MHz,Chloroform-d)δ7.45–7.37(m,2H),7.35–7.29(m,4H),7.21– 7.09(m,2H),5.71(s,2H),4.41–4.14(m,7H),4.05–3.93(m,2H),3.89–3.77(m,6H),3.75 –3.45(m),3.42–3.28(m,12H),3.13–2.82(m,6H),2.40–1.75(m,14H),1.69–1.50(m, 4H),1.29–1.06(m,4H),0.95(t,J＝7.3Hz,3H),0.90(t,J＝7.3Hz,3H)。

Example 57: synthesis of Compound S57

470mg (1.10mmol, see example 31 for synthesis method), 1008mg (0.50mmol) of dihydroxypolyethylene glycol (OH-PEG-OH, average molecular weight 2000), 248mg (1.20mmol) of Dicyclohexylcarbodiimide (DCC), 11mg (0.05mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) are dissolved in 10mL of dichloromethane at room temperature, and reacted for 12 hours at room temperature under the protection of nitrogen. And (3) carrying out suction filtration to remove insoluble substances, carrying out rotary evaporation on the filtrate to remove dichloromethane, adding 250mL of diethyl ether, stirring for 1 hour, filtering, collecting a filter cake, and carrying out vacuum drying to obtain a crude product. The crude product was dialyzed to give S57740 mg as a white solid.

¹H NMR(300MHz,Chloroform-d)δ7.47–7.37(m,2H),7.36–7.28(m,4H),7.22– 7.09(m,2H),5.72(s,2H),4.42–4.15(m,10H),3.87–3.77(m,6H),3.74–3.45(m),3.41– 3.27(m,12H),3.12–2.82(m,6H),2.40–1.72(m,14H),1.70–1.50(m,4H),1.35(d,J＝6.8 Hz,6H),1.29–1.07(m,4H),0.94(t,J＝7.3Hz,3H),0.91(t,J＝7.3Hz,3H)。

The compounds S58 to S65 in examples 58 to 65 below were all obtained by the synthesis method of example 57, and they were replaced with the corresponding amino acids and dihydroxy polyethylene glycols of different molecular weights.

Example 58: synthesis of Compound S58

¹H NMR(300MHz,Chloroform-d)δ7.46–7.37(m,2H),7.36–7.28(m,4H),7.22– 7.09(m,2H),5.72(s,2H),4.41–4.15(m,10H),3.87–3.77(m,6H),3.75–3.45(m),3.41– 3.27(m,12H),3.12–2.82(m,6H),2.39–1.73(m,14H),1.69–1.50(m,4H),1.35(d,J＝6.8 Hz,6H),1.29–1.07(m,4H),0.94(t,J＝7.3Hz,3H),0.91(t,J＝7.3Hz,3H)。

Example 59: synthesis of Compound S59

¹H NMR(300MHz,Chloroform-d)δ7.46–7.37(m,2H),7.36–7.28(m,4H),7.22– 7.09(m,2H),5.72(s,2H),4.42–4.15(m,10H),3.87–3.77(m,6H),3.75–3.44(m),3.40– 3.27(m,12H),3.13–2.82(m,6H),2.39–1.74(m,14H),1.69–1.50(m,4H),1.37–1.34(m, 6H),1.29–1.08(m,4H),0.94(t,J＝7.3Hz,3H),0.91(t,J＝7.3Hz,3H)。

Example 60: synthesis of Compound S60

¹H NMR(300MHz,Chloroform-d)δ7.48–7.37(m,2H),7.35–7.27(m,4H),7.23– 7.09(m,2H),5.72(s,2H),4.41–4.15(m,9H),3.87–3.77(m,6H),3.76–3.45(m),3.41– 3.27(m,12H),3.15–2.84(m,8H),2.40–1.73(m,14H),1.69–1.50(m,4H),1.36–1.33(m, 6H),1.28–1.06(m,4H),0.95(t,J＝7.3Hz,3H),0.92(t,J＝7.3Hz,3H)。

Example 61: synthesis of Compound S61

¹H NMR(300MHz,Chloroform-d)δ7.48–7.37(m,2H),7.36–7.27(m,4H),7.23– 7.09(m,2H),5.72(s,2H),4.42–4.15(m,9H),3.87–3.76(m,6H),3.77–3.45(m),3.42– 3.26(m,12H),3.14–2.84(m,8H),2.40–1.72(m,14H),1.69–1.50(m,4H),1.36–1.33(m, 6H),1.29–1.06(m,4H),0.95(t,J＝7.3Hz,3H),0.92(t,J＝7.3Hz,3H)。

Example 62: synthesis of Compound S62

¹H NMR(300MHz,Chloroform-d)δ7.51–7.38(m,2H),7.34–7.26(m,4H),7.24– 7.10(m,2H),5.69(s,2H),4.55–4.32(m,4H),4.26–4.01(m,4H),3.89–3.78(m,6H),3.75 –3.46(m),3.40–3.24(m,12H),3.20–2.79(m,6H),2.78–2.58(m,4H),2.38–1.75(m, 14H),1.69–1.50(m,4H),1.30–1.09(m,4H),0.96(t,J＝7.3Hz,3H),0.92(t,J＝7.3Hz, 3H)。

Example 63: synthesis of Compound S63

¹H NMR(300MHz,Chloroform-d)δ7.52–7.38(m,2H),7.34–7.26(m,4H),7.24– 7.11(m,2H),5.69(s,2H),4.56–4.32(m,4H),4.26–4.01(m,4H),3.89–3.79(m,6H),3.76 –3.46(m),3.41–3.24(m,12H),3.20–2.79(m,6H),2.78–2.57(m,4H),2.39–1.76(m, 14H),1.69–1.50(m,4H),1.31–1.09(m,4H),0.96(t,J＝7.3Hz,3H),0.92(t,J＝7.3Hz, 3H)。

Example 64: synthesis of Compound S64

¹H NMR(300MHz,Chloroform-d)δ7.51–7.38(m,2H),7.34–7.25(m,4H),7.24– 7.10(m,2H),5.69(s,2H),4.55–4.32(m,4H),4.27–4.01(m,4H),3.89–3.78(m,6H),3.76 –3.45(m),3.40–3.24(m,12H),3.21–2.79(m,6H),2.78–2.58(m,4H),2.39–1.74(m, 14H),1.70–1.51(m,4H),1.31–1.09(m,4H),0.96(t,J＝7.3Hz,3H),0.92(t,J＝7.3Hz, 3H)。

Example 65: synthesis of Compound S65

¹H NMR(300MHz,Chloroform-d)δ7.49–7.35(m,2H),7.32–7.23(m,4H),7.21– 7.07(m,2H),5.69(s,2H),4.29–4.06(m,4H),3.87–3.77(m,6H),3.75–3.46(m),3.39–3.21(m,12H),3.20–2.95(m,4H),2.92(s,6H),2.90–2.77(m,2H),2.62–2.55(m,4H),2.36 –1.71(m,14H),1.68–1.49(m,4H),1.29–1.07(m,4H),0.95(t,J＝7.4Hz,3H),0.90(t,J＝ 7.4Hz,3H)。

Effect example 1: vincamine derivative Vin3 for improving motor nerve conduction speed and sensory loss of diabetic peripheral neuropathy model mice

The invention takes a PEGylated vincamine derivative Vin3 as an example to illustrate the therapeutic effect in vivo, and the structure of Vin3 is shown in the following figure. The invention detects the influence of the longitramine derivative Vin3 on the motor nerve conduction velocity and pain response of a diabetic peripheral neuropathy model mouse on an STZ-induced type 1 diabetes mouse and a type 2 diabetes db/db mouse. The experimental result shows that the Vin3 can obviously improve the motor nerve conduction velocity and the sensory deletion phenomenon of the diabetic peripheral neuropathy model mouse.

1. Principle of experiment

The diabetic peripheral neuropathy firstly affects sensory neurons, and the main pathological change is axonal atrophy degeneration, even disappearance and the like. It is considered that the therapeutic effect against diabetic peripheral neuropathy can be achieved by protecting and promoting the growth of sensory neuron processes. In addition, diabetic peripheral neuropathy patients may develop symptoms such as decreased nerve conduction velocity, hypoesthesia, and asthenia. The behavioral indexes such as motor nerve conduction velocity, mechanical pain threshold value, thermal pain reaction time and the like of the mouse are detected on the diabetic peripheral neuropathy model mouse, and the behavioral indexes can be used for evaluating the improvement effect of the vincamine derivative Vin3 on the behavioral level of the diabetic peripheral neuropathy individual; the mouse peripheral neuron axon growth condition after administration is detected, and the protective effect of the compound Vin3 on the peripheral neuron axon can be evaluated.

2. Experimental materials and methods

1) Animal grouping administration

Type 1 diabetic mice: male 8-week-old C57BL/6J mice were purchased from Beijing Wittiulihua laboratory animals technical Limited. Carrying out intraperitoneal injection of STZ (150mg/kg) after one week, carrying out blood sampling on tail veins after one week to detect blood sugar, and selecting mice with random blood sugar of more than 16 mmol/L. The STZ was divided into groups 6 weeks after injection according to body weight and blood glucose, respectively: the control group (non-diabetic mice), the model group (DPN mice), and the Vin3 administration group (40mg/kg) were subjected to continuous gavage for 4 weeks.

Type 2 diabetic mice: male 18-week-old BKS db/db mice were purchased from Jiangsu Jiejiaokang Biotech Co., Ltd. After adaptive feeding for one week, grouping according to weight and blood sugar, respectively: the control group (db/m mice), the model group (db/db mice), and the Vin3 administration group (40mg/kg) were administered by continuous gavage for 4 weeks.

2) Motor nerve conduction velocity detection experiment

Before administration, Motor Neural Conduction Velocities (MNCVs) were measured on both left and right sides of each group of mice at 2 weeks and 4 weeks before administration, and the mean value of the velocities on both sides was used as the neural conduction velocity of each mouse. The specific experimental method comprises the following steps: placing electrodes: the stimulating electrode is arranged on the nerve trunk, the recording electrode is arranged on the muscle abdomen, and the reference electrode is arranged on the tendon; the ground line is disposed between the stimulating electrode and the recording electrode. Calculating motor nerve conduction velocity: the nerve conduction velocity is determined by that the distance between the far end and the near end is divided by the latency difference between the two points. The calculation formula is as follows: the nerve conduction velocity (m/s) is equal to the distance between two points (cm) × 10/latency difference between two points (ms).

3) Mechanical pain threshold detection experiment

This experiment is based on the principle that the rodent paw will retract in response to mechanical stimulation. The Von frey tactile measurement kit is purchased from Ugo Basile company, the Von frey fiber can provide 0.008g-300g of stimulation force, and the thickness of the fiber determines the stimulation force. The specific experimental method comprises the following steps: selecting the fiber filaments with proper thickness, vertically stimulating the skin, replacing the fiber filaments, and adjusting the stimulation force until the fiber filaments are bent. One mouse was assayed 6 times to determine the threshold value according to 50% threshold ═ 10^ (x)^f+ k δ))/10000 the mechanical pain threshold of 50% of mice was calculated and measured before and weekly after dosing, respectively.

4) Thermal pain latency detection experiment

Thermal pain sensitivity of the animals was characterized by measuring their response to infrared thermal stimuli using an infrared plantar pain tester (37370, available from Ugo Basile) before and weekly after administration. The method comprises the steps of putting a mouse into a single cage matched with a machine in advance to adapt for 20min, then carrying out thermal stimulation on the sole of the mouse, gradually raising the temperature along with the time, and automatically stopping the machine and recording the thermal stimulation time when the mouse lifts the foot or moves away from a heat source, wherein the thermal stimulation time is the thermal pain latent period. The thermal pain sensitivity was evaluated by comparing the length of the thermal stimulation time.

5) Dorsal root ganglion neuron cell axon growth experiment

After the administration, the neurons of the fresh Dorsal Root Ganglion (DRG) of the mice are digested into single cell suspension, and then inoculated into a culture plate for adherent culture overnight. Beta-tubulin III (Sigma) protein of DRG neuron primary cells was labeled by immunofluorescence staining. Fluorescence microscopy (Leica) imaged, and then the total length of axons of individual DRG neuron cells was followed with Neuro J plug-in Image J software.

3. Results of the experiment

As shown in tables 1-3, the vincamine derivative Vin3 can significantly improve motor nerve conduction velocity and sensory loss symptoms of type 1 diabetic peripheral neuropathy mice. All experimental data for this work are shown as Mean ± SD., and all histograms and graphs are plotted and analyzed using GraphPad Prism 8 software. Significance analysis was performed between the two sets of data using T test.^*p<0.05，^**p<0.01，^***p<0.001 represents model group vs. control group;^#p<0.05，^##p<0.01， ^###p<0.001 represents the administration group vs. the model group.

TABLE 1 motor nerve conduction velocity (Mean + -SD., n-6, m/s)

TABLE 250% mechanical pain threshold (Mean + -SD., n-7, g)

TABLE 3 incubation period for thermal pain sensation (Mean + -SD., n-7, s)

The results are shown in tables 4-6, and the vincamine derivative Vin3 can obviously improve motor nerve conduction speed and sensory loss symptoms of type 2 diabetic mice. Significance analysis was performed between the two sets of data using T test.^*p<0.05，^**p<0.01， ^***p<0.001 represents db/dbvs. db/m;^#p<0.05，^##p<0.01，^###p<0.001 represents Vin3 processed vs. db/db.

TABLE 4 motor nerve conduction velocity (Mean + -SD., n-6, m/s)

TABLE 550% mechanical pain threshold (Mean + -SD., n-7, g)

TABLE 6 incubation period for thermal pain sensation (Mean + -SD., n-7, s)

The result is shown in figure 1, the vincamine derivative Vin3 can obviously improve the axon growth condition (fluorescence and quantitative) of DRG neurons in a type 1 diabetic peripheral neuropathy model mouse. The statistics adopts T-test, n is 5,^*p<0.05， ^**p<0.01，^***p<0.001, respectively, compared to the model groups. As shown in figure 2, the vincamine derivative Vin3 also can obviously promote the axonal growth of DRG neurons in a type 2 diabetes peripheral neuropathy model mouse (a fluorescence graph and a quantitative graph). The statistics adopts T-test, n is 7,^*p<0.05，^**p<0.01，^***p<0.001, respectively, compared to the model groups. The longitramine derivative Vin3 is shown to have protective effect on peripheral sensory neuron axons.

Effect example 2: vincamine derivative Vin3 for promoting foot ulcer wound healing of diabetic foot model rat

The invention detects the influence of the vincamine derivative Vin3 on the healing of the ulcer wound of the foot of the diabetic foot model rat on the diabetic foot model rat body induced by STZ. The experimental result shows that after the foot ulcer of the diabetic rat, the compound Vin3 is used for treatment, so that the healing of the foot ulcer wound can be obviously promoted, and the injury area is reduced.

1. Principle of experiment

Diabetic Foot Ulcers (DFUs) are a common manifestation of Diabetic feet and are also a major cause of amputation in Diabetic patients. In the experiment, wounds with a certain area are formed on the dorsum of the foot of a diabetic rat at one time to form a diabetic foot ulcer model, and then a compound Vin3 is given for treatment. The effect of the vincamine derivative Vin3 on wound healing of diabetic foot ulcers was assessed by photographing the wounds and quantifying their area size.

2. Experimental materials and methods

1) Establishment of diabetic rat model

Male 5 week old SPF grade Sprague Dawley rats were purchased from Experimental animals technology, Inc. of Wintolite, Beijing. Carrying out intraperitoneal injection of STZ (100mg/kg) after one week, carrying out tail vein blood sampling after 72h to detect blood sugar, and selecting mice with random blood sugar of more than 16.7 mmol/L.

2) Establishment of diabetic foot ulcer rat model

4 weeks after STZ injection using an inhalation anesthesia machine, isoflurane inhalation induced anesthesia, a complete circular skin wound was created on the hind instep of the diabetic rat by a disposable 5mm biopsy punch and Westcott scissors. After the diabetic foot ulcer rat model is established, the model is divided into a control group (non-diabetic rat), a model group (DFU rat) and a Vin3 administration group (40mg/kg) according to blood sugar and weight indexes, and the administration is carried out for 4 weeks by continuous intragastric administration.

3) Diabetic foot rat ulcer area observation

The size of the wound area of the foot ulcer in each mouse was observed on day 0, day 3, day 7, day 14, day 21 and day 28 after the administration, respectively. The area of the lesion on the foot of each rat was measured with a ruler, and after images were taken with a camera, the area of the wound was quantified by conversion to a scale using Image J software.

3. Results of the experiment

As shown in fig. 3, the vincamine derivative Vin3 can obviously promote the healing of diabetic foot ulcer wounds. The wound area quantification results are shown in table 7, and significance analysis was performed between the two sets of data using T test.^*p<0.05，^**p<0.01，^***p<0.001 represents model group vs. control group;^#p<0.05，^##p<0.01，^###p<0.001 represents the administration group vs. the model group.

Table 7 foot ulcer wound area (Mean ± SD., n-7, mm)²)

Example 3: vinblastine derivative Vin3 for relieving bleomycin-induced pulmonary fibrosis degree of mice

The invention utilizes Micro CT to detect the protective effect of the vincamine derivative Vin3 on the pulmonary fibrosis tissues of mice on bleomycin-induced pulmonary fibrosis model mice. The experimental result shows that the treatment of the compound Vin3 can obviously reduce the pulmonary fibrosis degree of the mice.

1. Principle of experiment

The experiment resulted in a pulmonary fibrosis model by oral tracheal instillation of bleomycin solution in mice, followed by treatment with compound Vin 3. The influence of the vincamine derivative Vin3 on the degree of bleomycin-induced pulmonary fibrosis of mice was evaluated by CT photographing and three-dimensional reconstruction of the mouse lungs and quantifying the volume of normal tissues of the mouse lungs.

2. Experimental materials and methods

Male 7-week-old C57BL/6J mice were purchased from Beijing Wittingle, and after one week of acclimatization, lung development of the mice was examined by micro-CT. Mice were divided into a blank control group (normal mice), a model group and a Vin3 administration group (100mg/kg) according to the size of the lung normal tissue volume. Then, a suction type anesthesia machine is adopted, isoflurane is inhaled to induce anesthesia, 5mg/kg of bleomycin solution is instilled into mice of a model group and an administration group through an oral trachea, a pulmonary fibrosis model is established, and an equal volume of bleomycin solvent is instilled into a control group. Administration was started the next day and continued by gavage for 20 days. Before molding, micro-CT imaging experiments are carried out on the 10 th day and the 20 th day after molding, and the pulmonary fibrosis condition of the mouse is observed.

3. Results of the experiment

As shown in figure 4, the vincamine derivative Vin3 can obviously reduce the degree of pulmonary fibrosis of mice, and has a certain protective effect on lung tissues. After 20 days of administration, the lung normal tissue volume of the mice in the administration group was significantly larger than that in the model group. The quantification of the normal tissue volume of the mouse lung is shown in table 8, and significance analysis was performed between the two data sets using T test.^*p<0.05，^**p<0.01，^***p<0.001 represents model group vs. control group;^#p<0.05，^##p<0.01，^###p<0.001 represents the administration group vs. model group.

Table 8 lung normal tissue volume (Mean ± SD., n-5, mm)³)

The above embodiments are only exemplary embodiments of the present invention, and are not intended to limit the present invention, and the protection scope of the present invention is defined by the claims. Various modifications and equivalents may be made by those skilled in the art within the spirit and scope of the present invention, and such modifications and equivalents should also be considered as falling within the scope of the present invention.

Claims (12)

1. A vincamine PEG derivative is characterized by comprising compounds shown in formulas I and II, pharmaceutically acceptable salts, metabolic precursors, metabolites or isomers thereof, wherein the structural formula of the compounds is shown as follows:

in formula I and formula II:

l is a single bond,

Amino acids, or, di-hexapeptides formed by amino acid condensation;

x and Y are independently selected from the group consisting of a single bond, a,

Amino acids, di-hexapeptides formed by amino acid condensation;

t is an integer of 1 to 6;

R¹is hydrogen, unsubstituted or R^1-1Substituted C_1-6Alkyl radical, C_6-10Aryl radical, C_3-10Cycloalkyl, one or more than one heteroatom selected from N, O, S, "5-10 membered heteroaryl with 1-3 heteroatoms," one or more than one heteroatom selected from N, O, S, "4-10 membered heterocycloalkyl with 1-3 heteroatoms," C_3-10Cycloalkyl- (C)_1-4Alkyl) -, or, C_6-12Aryl radical- (C)_1-4Alkyl) -;

R^1-1is halogen, hydroxy, amino or C_1-4An alkyl group;

n is an integer of 0 to 500;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-polyethylene glycol of linear, dendritic, star-shaped or hyperbranched structure;

r is-O (R)²)、-NH(R³)、-COOH；

R²Is hydrogen or C_1-6An alkyl group;

R³is hydrogen or C_1-6An alkyl group.

2. A vincamine PEG derivative according to claim 1, wherein t is an integer from 1 to 3;

and/or, when L is an amino acid, said amino acid is alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, or histidine;

and/or, when L is a di-hexapeptide formed by condensation of amino acids, said amino acids are the same or different;

and/or, when L is a di-hexapeptide formed by condensation of amino acids, the di-hexapeptide is a di-tetrapeptide;

and/or, when X and Y are independently selected from amino acids, said amino acids are alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, or histidine;

and/or, when X and Y are independently selected from the group consisting of di-hexapeptides formed by the condensation of amino acids, said amino acids are the same or different;

and/or, when X and Y are independently selected from the group consisting of a di-hexapeptide formed by the condensation of amino acids, said di-hexapeptide is a di-tetrapeptide;

and/or when R¹Is unsubstituted or R^1-1Substituted C_1-6When it is alkyl, said R^1-1Is one or more, when there are more than one R^1-1When R is said^1-1May be the same or different;

and/or when R¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R¹Is C_6-10When aryl, said C_6-10Aryl is phenyl;

and/or when R¹Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl being C_3-6A cycloalkyl group;

and/or when R¹Is C_6-12Aryl radical- (C)_1-4Alkyl) -said C_6-12Aryl radical- (C)_1-4Alkyl) -is phenyl- (C)_1-2Alkyl) -;

and/or n is an integer of 0-300;

and/or the presence of a gas in the gas,

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-a linear polyethylene glycol;

and/or when R²Is C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R³Is C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group.

3. Vincamine PEG derivative according to claim 1, and/or, when L is an amino acid, the amino acid is alanine, phenylalanine, methionine, glutamic acid, aspartic acid, valine, leucine and proline, preferably alanine, phenylalanine, valine, leucine or serine;

and/or, when L is a di-hexapeptide formed by condensation of amino acids, said amino acids are glycine, alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine or histidine, preferably glycine, alanine, valine, serine or phenylalanine;

and/or, when X and Y are independently selected from the group consisting of a di-hexapeptide formed by the condensation of amino acids, said di-hexapeptide is glycine-, glycine-alanine, alanine-alanine, glycine-valine, glycine-serine, or glycine-phenylalanine;

and/or, when X and Y are independently selected from amino acids, said amino acids are alanine, phenylalanine, methionine, glutamic acid, aspartic acid, leucine and proline, preferably alanine, phenylalanine, valine, leucine or serine;

and/or, when X and Y are independently selected from the group consisting of a di-hexapeptide formed by the condensation of amino acids, said amino acid is glycine, alanine, valine, leucine, isoleucine, phenylalanine, proline, tryptophan, serine, tyrosine, cysteine, methionine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine or histidine, preferably glycine, alanine, valine, serine or phenylalanine;

and/or, when X and Y are independently selected from the group consisting of a di-hexapeptide formed by the condensation of amino acids, said di-hexapeptide is glycine-, glycine-alanine, alanine-alanine, glycine-valine, glycine-serine, or glycine-phenylalanine;

and/or when R¹Is unsubstituted or R^1-1Substituted C_1-6When it is alkyl, said R^1-1The number of (a) is 1,2 or 3;

and/or when R¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl or isopropyl;

and/or when R¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl or isopropyl;

and/or when R¹Is unsubstituted or R^1-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isobutyl or isopropyl;

and/or when R¹Is unsubstituted or R^1-1Substituted C_1-6When it is alkyl, said R^1-1Substituted C_1-6Alkyl is

And/or when R¹Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

and/or when R¹Is C_6-12Aryl radical- (C)_1-4Alkyl) -said C_6-12Aryl radical- (C)_1-4Alkyl) -is phenyl- (C)_1-2Alkyl groups);

and/or when R¹Is C_6-12Aryl radical- (C)_1-4Alkyl) -said C_6-12Aryl radical- (C)_1-4Alkyl) -is benzyl or phenethyl;

and/or n is an integer of 0-200;

and/or when R²Is C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isopropyl or isobutyl;

and/or when R³Is C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isopropyl or isobutyl.

4. A vincamine PEG derivative according to claim 1, wherein L is

Amino acids or amino acids condensed to form di-hexapeptides;

and/or X is

Amino acids or amino acids condensed to form di-hexapeptides;

and/or Y is

Amino acids or amino acids condensed to form di-hexapeptides;

and/or, t is 1 or 2;

and/or, R¹Is hydrogen, unsubstituted or R^1-1Substituted C_1-6Alkyl radical, C_6-10Aryl or C_3-10A cycloalkyl group;

and/or, R^1-1Is a hydroxyl group;

and/or n is an integer of 1-200;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-a linear polyethylene glycol;

and/or R is-O (R)²)；

And/or, R²Is hydrogen or C_1-6An alkyl group.

5. A vincamine PEG derivative according to claim 1, wherein L is

Amino acids, or, di-hexapeptides formed by amino acid condensation;

x is

Amino acids or amino acids condensed to form di-hexapeptides;

y is

Amino acids or amino acids condensed to form di-hexapeptides;

t is 1 or 2;

R¹is hydrogen, unsubstituted or R^1-1Substituted C_1-6Alkyl radical, C_6-10Aryl or C_3-10A cycloalkyl group;

R^1-1is a hydroxyl group;

n is an integer of 0 to 200;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-a linear polyethylene glycol;

r is-O (R)²)；

R²Is hydrogen or C_1-6An alkyl group.

6. A vincamine PEG derivative according to claim 1, wherein L is

Amino acids or amino acids condensed to form di-hexapeptides;

x is

Amino acids or amino acids condensed to form di-hexapeptides;

y is

Amino acids or amino acids condensed to form di-hexapeptides;

t is 1 or 2;

R¹is hydrogen, unsubstituted or R^1-1Substituted C_1-6Alkyl or C_3-10A cycloalkyl group;

R^1-1is a hydroxyl group;

n is an integer of 0 to 100;

is prepared by reacting a compound having a structure containing- (OCH)₂CH₂)_n-a linear polyethylene glycol;

r is-O (R)²)；

R²Is hydrogen or C_1-6An alkyl group.

7. A vincamine PEG derivative according to claim 1, wherein the compound of formula i is any one of the following:

8. a process for the preparation of a compound of formula i as claimed in claim 1, the synthesis of which comprises the steps of: in a solvent, under the action of alkali and a condensing agent, carrying out a condensation reaction shown in the specification on a compound shown in a formula III and a compound shown in a formula IV;

wherein L and R are as defined in any one of claims 1 to 6.

9. A process for the preparation of a compound of formula ii as claimed in claim 1, the synthesis of which comprises the steps of: in a solvent, under the action of alkali and a condensing agent, carrying out a condensation reaction as shown in the specification on a compound shown in a formula V and a compound shown in a formula III;

wherein X and Y are as defined in any one of claims 1 to 6.

10. A pharmaceutical composition comprising a compound of formula i and formula ii, pharmaceutically acceptable salts thereof, metabolic precursors thereof, metabolites thereof or isomers thereof, as claimed in any one of claims 1 to 7, and a pharmaceutically acceptable adjuvant.

11. Use of a compound of formula i and formula ii, pharmaceutically acceptable salts thereof, metabolic precursors thereof, metabolites thereof or isomers thereof, as claimed in any one of claims 1 to 7, in the manufacture of a medicament for the prevention or treatment of diabetic complications; the diabetes mellitus is type 1 diabetes mellitus and type 2 diabetes mellitus.

12. Use of a compound of formula i and formula ii, pharmaceutically acceptable salts thereof, metabolic precursors thereof, metabolites thereof or isomers thereof, as claimed in any one of claims 1 to 7, in the manufacture of a medicament for the prophylaxis or treatment of pulmonary fibrosis.

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