CN114249721B

CN114249721B - Application of vincamine PEG derivative in preparing medicament for treating diabetic peripheral neuropathy, diabetic foot and pulmonary fibrosis

Info

Publication number: CN114249721B
Application number: CN202111432744.7A
Authority: CN
Inventors: 沈旭; 蒋晟; 王佳颖; 张阔军; 黄玉洁; 张文波; 许瑞; 卢之瑀
Original assignee: China Pharmaceutical University; Nanjing University of Chinese Medicine
Current assignee: China Pharmaceutical University; Nanjing University of Chinese Medicine
Priority date: 2021-11-29
Filing date: 2021-11-29
Publication date: 2023-10-17
Anticipated expiration: 2041-11-29
Also published as: CN114249721A

Abstract

The invention relates to a PEGylated vincamine derivative and application thereof. The invention specifically discloses a PEGylated vincamine derivative shown in formulas I and II, and pharmaceutically acceptable salts, metabolic precursors, metabolites or isomers thereof. Experiments show that the vincamine derivative can promote the growth of peripheral sensory neuron protrusion, improve the nerve conduction speed and sensory deficit symptoms of a diabetic mouse, and promote the healing of foot ulcer wounds of the diabetic mouse, so that the vincamine derivative has good treatment effect on peripheral neuropathy of diabetic complications and diabetic feet. Experiments show that the vincamine derivative can also obviously reduce the pulmonary fibrosis degree of mice induced by bleomycin and has a protective effect on pulmonary tissues.

Description

Application of vincamine PEG derivative in preparing medicament for treating diabetic peripheral neuropathy, diabetic foot and pulmonary fibrosis

Technical Field

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

Background

Diabetes is a serious worldwide disease. It is estimated that by 2045 years, the number of diabetics will reach 6.29 billion. Diabetes now becomes a third non-infectious disease that threatens human health and life following cardiovascular disease and tumors. In China, diabetes has changed from rare diseases to epidemic diseases, and the prevalence rate has risen from 0.67% on average to 11.6% in recent 30 years. Diabetes patients are easily afflicted with various diseases complicated with diabetes, including diabetic neuropathy, nephropathy, diabetic foot, etc., without effective control and treatment methods, and binocular blindness, renal failure, even limb disability and death occur.

Diabetic peripheral neuropathy (Diabetic peripheral neuropathy, DPN) is a chronic neuropathy of diabetes caused by long-term hyperglycemia, with a incidence rate of up to 50% -80%, which is the most common chronic complication of diabetes. Diabetic peripheral neuropathy can involve motor, sensory and autonomic nerves, creating motor and sensory disorders. Clinically, the vibration and thermal perception thresholds of diabetic peripheral neuropathy patients increase, and these thresholds can develop sensory loss with degeneration of all fibers in the peripheral nerve. Currently, the diagnosis method of diabetic peripheral neuropathy mainly comprises sensory detection, electrophysiological detection, nerve fiber morphological examination, imaging examination and the like. However, the clinical manifestations of diabetic peripheral neuropathy are various, the classification methods at home and abroad are different, the screening and inspection methods are various, the diagnosis standards are different, and the prevention and treatment of the diabetic peripheral neuropathy are all clinical difficulties. Although scientists have made breakthrough progress in the study of diabetic peripheral neuropathy with rapid development of science and technology, there is still a lack of specific drugs for radical treatment of diabetic peripheral neuropathy. Thus, there is no delay in the research and development of novel therapeutic strategies and novel compounds.

Diabetic Foot (DF) is a disorder that causes Diabetic vascular lesions and/or neuropathy and infection due to long-term hyperglycemia, thereby causing necrosis of the tissues of the foot or lower limb of a Diabetic patient. Diabetic foot is one of the most serious complications of diabetes, is one of the main causes of disability and death for diabetics, and is also a great public health problem causing a heavy burden to society. With the increasing incidence of diabetes, the number of individuals with diabetic foot disease increases year by year. The incidence rate of diabetic foot of diabetics over 50 years old in China is up to 8.1 percent. Unfortunately, the prognosis for diabetic foot is poor, even higher than the mortality and disability rate for most cancers (excluding lung cancer, pancreatic cancer, etc.). And the diabetic foot treatment cost is high, long-term treatment is required, foot ulcers of patients can be healed mainly through medical care in clinic, and the risk of amputation is avoided. However, these treatments do not completely radically cure the diabetic foot and are prone to relapse. Therefore, the research on pathogenesis of the diabetic foot and the development of novel and effective therapeutic drugs for the diabetic foot have very important practical significance.

Pulmonary fibrosis (Pulmonary fibrosis, PF) is a chronic, progressive, fatal pulmonary disease, an end-stage of a variety of diseases, the pathogenesis of which has not been elucidated to date. Pulmonary fibrosis severely affects the respiratory function of the human body, manifesting as dry cough, progressive dyspnea (inadequate consciousness), and with increased disease and lung injury, patient respiratory function is continually worsened. The incidence and mortality of idiopathic pulmonary fibrosis increases year by year, with an average survival of only 2.8 years after diagnosis, and mortality higher than most tumors, known as a "neoplastic disease". At present, pirfenidone, nilamide, oxygen therapy, invasive/noninvasive mechanical ventilation and lung transplantation are mostly adopted for treating patients with pulmonary fibrosis. However, these methods have high cost, insignificant efficacy, insufficient transplantation donors or immunological rejection, and poor medical compliance of patients, which limits their wide application. Therefore, it is important to actively explore the pathogenesis of pulmonary fibrosis and to develop new intervention drugs.

Vincamine (Vincamine) is a monoterpene indole alkaloid found in motor gas lengthened spring flowers. 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 a nootropic function. However, as with many natural products, vincamine also suffers from poor water solubility, unstable metabolism, short duration of action, etc. From previous work, vincamine has the potential to treat diabetic complications and pulmonary fibrosis. Therefore, the search for novel vincamine derivatives is of great importance for the treatment of diabetic complications and pulmonary fibrosis.

Polyethylene glycol (Polyethylene glycol, PEG) is a type of water-soluble polyether with low molecular weight obtained by gradually carrying out addition polymerization on ethylene oxide and water or ethylene glycol. The low molecular weight polyethylene glycol is colorless, odorless and sticky liquid with hygroscopicity, and has ether chains and hydroxyl groups in molecules, so that the low molecular weight polyethylene glycol has unique solubility and good biocompatibility, and has important application prospects in the fields of medicines, materials, engineering and the like.

Disclosure of Invention

The invention aims to solve the technical problem that PEG is connected with vincamine by utilizing the characteristics of innocuity and easy combination, and overcomes the characteristic that the vincamine is insoluble in water. The invention provides a vincamine derivative, a preparation method, a pharmaceutical composition and application thereof. The PEGylated vincamine derivative has stable structure and good water solubility, and has good treatment effect on diabetic peripheral neuropathy, diabetic patients and pulmonary fibrosis.

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

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

in formula I and formula II:

l is a single bond,Amino acids, or di-hexapeptides formed by condensation of amino acids;

x and Y are independently selected from single bond,Amino acids, or di-hexapeptides formed by condensation of amino acids;

t is an integer from 1 to 6;

R ¹ is hydrogen, unsubstituted or R ^1-1 Substituted C _1-6 Alkyl, C _6-10 Aryl, C _3-10 Cycloalkyl, one or more of the "hetero atom is selected from N, O, S," 5-10 membered heteroaryl group with 1-3 hetero atoms, or one or more of the "hetero atom is selected from N, O, S," 4-10 membered heterocycloalkyl group with 1-3 hetero atoms, C _3-10 Cycloalkyl- (C) _1-4 Alkyl) -, or C _6-12 Aryl- (C) _1-4 Alkyl) -;

R ^1-1 halogen, hydroxy, amino or C _1-4 An alkyl group;

n is an integer of 0 to 500;

is provided with a compound containing- (OCH) ₂ CH ₂ ) _n -polyethylene glycols of linear, tree, star or hyperbranched structure;

r is-O (R) ² )、-NH(R ³ ) -COOH;

R ² is hydrogen or C _1-6 An alkyl group;

R ³ is hydrogen or C _1-6 An 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 amino acid condensation, the di-hexapeptide is a di-tetrapeptide;

when X and Y are independently selected from amino acids, the 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 di-hexapeptides formed by the condensation of amino acids, the 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 is ¹ Is unsubstituted or R ^1-1 Substituted C _1-6 In the case of alkyl, said R ^1-1 Is one or more, when there are a plurality of R ^1-1 When said R is ^1-1 May be the same or different;

when R is ¹ Is unsubstituted or substitutedR ^1-1 Substituted C _1-6 In the case of alkyl, said C _1-6 Alkyl is C _1-4 An alkyl group;

when R is ¹ Is C _6-10 Aryl, the C _6-10 Aryl is phenyl;

when R is ¹ Is C _3-10 In the case of cycloalkyl, said C _3-10 Cycloalkyl radicals are C _3-6 Cycloalkyl;

when R is ¹ Is C _6-12 Aryl- (C) _1-4 Alkyl) -at the time of said C _6-12 Aryl- (C) _1-4 Alkyl) -being phenyl- (C _1-2 Alkyl) -;

n is an integer of 0 to 300;

is provided with a compound containing- (OCH) ₂ CH ₂ ) _n -a linear polyethylene glycol;

when R is ² Is C _1-6 In the case of alkyl, said C _1-6 Alkyl is C _1-4 An alkyl group;

when R is ³ Is C _1-6 In the case of alkyl, said C _1-6 Alkyl is C _1-4 An 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 condensation of 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 amino acid condensed di-hexapeptides, said di-hexapeptides are glycine-, glycine-alanine, alanine-alanine, glycine-valine, glycine-serine, or glycine-phenylalanine;

When X and Y are independently selected from the group consisting of 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 condensation of 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 R is ¹ Is unsubstituted or R ^1-1 Substituted C _1-6 In the case of alkyl, said R ^1-1 Is 1, 2 or 3;

when R is ¹ Is unsubstituted or R ^1-1 Substituted C _1-6 In the case of alkyl, said C _1-6 Alkyl is methyl, ethyl, propyl or isopropyl, preferably methyl, ethyl or propyl;

When R is ¹ Is unsubstituted or R ^1-1 Substituted C _1-6 In the case of alkyl, said C _1-6 Alkyl is methyl, ethyl, propyl or isopropyl, preferably methyl;

when R is ¹ Is unsubstituted or R ^1-1 Substituted C _1-6 In the case of alkyl, said C _1-6 Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isobutyl or isopropyl;

when R is ¹ Is unsubstituted or R ^1-1 Substituted C _1-6 In the case of alkyl, said R ^1-1 Substituted C _1-6 Alkyl is

When R is ¹ Is C _3-10 In the case of cycloalkyl, said C _3-10 Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl;

when R is ¹ Is C _6-12 Aryl- (C) _1-4 Alkyl) -at the time of said C _6-12 Aryl- (C) _1-4 Alkyl) -being phenyl- (C _1-2 An alkyl group);

when R is ¹ Is C _6-12 Aryl- (C) _1-4 Alkyl) -at the time of said C _6-12 Aryl- (C) _1-4 Alkyl) -is benzyl or phenethyl;

n is an integer of 0 to 200;

when R is ² Is C _1-6 In the case of alkyl, said C _1-6 Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isopropyl or isobutyl, preferably methyl or ethyl;

when R is ³ Is C _1-6 In the case of alkyl, said C _1-6 Alkyl is methyl, ethyl, propyl, butyl, sec-butyl, isopropyl or isobutyl, preferably methyl or ethyl.

In one embodiment, certain groups of the compounds of formulas I and II are defined as follows (undefined groups are described in any of the previous embodiments):

L isAmino acids, or di-hexapeptides formed by condensation of amino acids;

x isAmino acids or di-hexapeptides formed by amino acid condensation;

y isAmino acids or di-hexapeptides formed by amino acid condensation;

t is 1 or 2;

R ¹ is hydrogen, unsubstituted or R ^1-1 Substituted C _1-6 Alkyl, C _6-10 Aryl or C _3-10 Cycloalkyl;

R ^1-1 is hydroxyl;

n is an integer of 0 to 200;

r is-O (R) ² )；

R ² Is hydrogen or C _1-6 An alkyl group.

l isAmino acids or di-hexapeptides formed by amino acid condensation;

x isAmino acids or di-hexapeptides formed by amino acid condensation;

y isAmino acids or di-hexapeptides formed by amino acid condensation;

t is 1 or 2;

R ¹ is hydrogen, unsubstituted or R ^1-1 Substituted C _1-6 Alkyl or C _3-10 Cycloalkyl;

R ^1-1 is hydroxyl;

n is an integer of 0 to 100;

is provided with a containing-(OCH ₂ CH ₂ ) _n -a linear polyethylene glycol;

r is-O (R) ² )；

R ² Is hydrogen or C _1-6 An alkyl group.

in one embodiment, the compounds of formulas I and II may have any of the following structures:

The invention also provides a preparation method of the compound shown in the formula I, and the synthesis method comprises the following steps: in a solvent, under the action of alkali and a condensing agent, performing condensation reaction between a compound shown in a formula III and a compound shown in a formula IV;

wherein L and R are as defined above.

The invention also provides a preparation method of the compound shown in the formula I, which comprises the following steps: in a solvent, under the action of alkali and condensing agent, performing condensation reaction between 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 in the formula I, pharmaceutically acceptable salt thereof, metabolic precursor thereof, metabolic product thereof or isomer thereof in preparing medicaments, wherein 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, a pharmaceutically acceptable salt thereof, a metabolic precursor thereof, a metabolite thereof, or an isomer thereof, as defined in any one of claims 1 to 7, and a pharmaceutical adjuvant.

In the pharmaceutical composition, the compound shown in the formula I, pharmaceutically acceptable salt, metabolic precursor, metabolite or isomer thereof can be used in an amount effective for treatment.

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

The pharmaceutical excipients can be those which are widely used in the field of pharmaceutical production. Adjuvants are used primarily to provide a safe, stable and functional pharmaceutical composition, and may also provide means for allowing the subject to dissolve at a desired rate after administration, or for promoting effective absorption of the active ingredient after administration of the composition. The pharmaceutical excipients may be inert fillers or provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients can comprise one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, sizing agents, disintegrants, lubricants, anti-adherents, glidants, wetting agents, gelling agents, absorption retarders, dissolution inhibitors, enhancing agents, adsorbents, buffering agents, chelating agents, preservatives, colorants, flavoring agents, and sweeteners.

The pharmaceutical compositions of the present invention may be prepared in accordance with the disclosure using any method known to those of skill 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, implantation, subcutaneous, intravenous, intra-arterial, intramuscular). The pharmaceutical compositions of the invention may also be in controlled or delayed release dosage forms (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, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serum formulations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry formulations which may 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; a liquid dosage form suitable for parenteral administration; suppositories and lozenges.

The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention prepared from the compounds of the present invention which have the specified substituents found herein with relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts may be obtained by contacting neutral forms of such compounds with a sufficient amount of a base in pure 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 the compounds of the present invention contain relatively basic functional groups, the acid addition salts may be obtained by contacting the neutral form of such compounds with a sufficient amount of an acid in pure solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, and the like; and organic acid salts including acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; also included are salts of amino acids such as arginine and the like, and salts of organic acids such as glucuronic acid and the like (see Berge et al, "Pharmaceutical Salts", journal of Pharmaceutical Science66:1-19 (1977)). Certain specific compounds of the invention contain basic and acidic functionalities that can be converted to either base or acid addition salts. Preferably, the salt is contacted with a base or acid in a conventional manner to isolate the parent compound, thereby regenerating the neutral form of the compound. The parent form of a 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, the preparation of such salts is as follows: prepared via reaction of 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 both. 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 (glucorination), enzymatic cleavage, etc. of the administered compound. Accordingly, the present invention includes metabolites of the compounds of the present invention, including compounds produced by a method of contacting a compound of the present invention with a mammal for a period of time sufficient to obtain the metabolites thereof.

Identification of metabolites is typically accomplished by preparing radiolabeled (e.g., 14C or ³ H) Isotopes, which are parenterally administered to animals, such as rats, mice, guinea pigs, monkeys, or humans, at a detectable dose (e.g., greater than about 0.5 mg/kg), allow sufficient time for metabolism to occur (typically about 30 seconds to 30 hours) and isolation of their conversion products from urine, blood, or other biological samples. These products are easy to isolate because they are labeled (others are isolated by using antibodies that are capable of binding to epitopes present in the metabolite). The metabolite structures are determined in a conventional manner, for example by MS, LC/MS or NMR analysis. In general, the analysis of metabolites is performed in the same manner as conventional drug metabolism studies known to those skilled in the art. So long as the metabolite products are not otherwise undetectable in vivo, they are useful in assays for therapeutic dosing of the compounds of the invention. The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more of the atoms comprising the compounds. For example, compounds can be labeled with radioisotopes, such as tritium @, for example ³ H) Iodine-125% ¹²⁵ I) Or C-14% ¹⁴ C) A. The application relates to a method for producing a fibre-reinforced plastic composite All isotopic variations of the compounds of the present application, whether radioactive or not, are intended to be encompassed within the scope of the present application.

The term "isomer" refers to two or more compounds that have the same molecular composition but different structures and properties.

It will be appreciated by those skilled in the art that, in accordance with the convention used in the art, the term "used herein to describe the structural formula of a group means that the corresponding group is attached to other fragments, groups, of the compounds of formulae I and II at that point.

"substitution" in the present application may be one or more, and when there are plural "substitution", the "substitution" may be the same or different.

The term "plurality" refers to 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, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl and the like.

The term "alkoxy" refers to the group-O-R ^Y Wherein R is ^Y Are alkyl groups 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 may be a bicyclic or tricyclic cycloalkyl having 4 to 10 carbon atoms.

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

The term "aryl" refers to an aryl group having the indicated number of carbon atomsA perfume group, preferably a monocyclic, bicyclic or tricyclic aromatic group, and when bicyclic or tricyclic, each ring satisfies the shock rule. C of the invention _6-10 Aryl of (a) refers to an aromatic group containing 6 to 10 carbon atoms, such as phenyl or naphthyl.

The term "heteroaryl" refers to an aromatic group containing heteroatoms, preferably an aromatic 5-6 membered monocyclic ring or a 9-10 membered bicyclic ring containing 1,2 or 3 atoms 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-oxadiazolyl, 1,2, 5-oxadiazolyl, 1,3, 4-oxadiazolyl, thiadiazolyl, dithiazolyl, tetrazolyl, pyridyl, pyranyl, thiopyranyl, 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, benzisozolyl, quinolinyl, isoquinolinyl.

The above preferred conditions can be arbitrarily combined on the basis of not deviating from the common knowledge in the art, and thus, each preferred embodiment of the present invention can be obtained.

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

The invention has the positive progress effects that:

(1) PEG modified vincamine has enhanced drug water solubility;

(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 graph and a quantitative histogram of vincamine derivative Vin3 capable of significantly promoting the growth of peripheral sensory neuron axons of type 1 diabetic mice.

Fig. 2 is an immunofluorescence and quantitative bar graph 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 vincamine derivative Vin3 can significantly promote healing of foot ulcer wound of diabetic foot rat.

Fig. 4 is a CT image of the vincamine derivative Vin3 capable of significantly improving the pulmonary fibrosis degree of mice.

Detailed Description

The invention is further illustrated below in connection with specific examples which should not be construed as limiting the invention.

Example 1: synthesis of Compound S1

Step one: synthesis of vincamine demethyl ester product

Vincamine 1.0g (2.8 mmol) was dissolved in 10mL of NaOH solution (1 mol/L) at room temperature and stirred, the solution gradually became clear from cloudiness, and the progress of the reaction was checked by TLC plate. After the vincamine reaction is finished, the reaction solution is placed at the temperature of 0 ℃ and the pH value is regulated to 3 by dilute hydrochloric acid, then methanol is removed by rotary evaporation, the filtration is carried out, a filter cake is collected, and the vacuum drying is carried out, thus obtaining a vincamine methyl-removing ester crude product of 0.919, and the crude product is directly put into the next step.

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

¹ H 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 (0.50 mmol) of vincamine glycinate, 61mg (0.50 mmol) of diethylene glycol monomethyl ether, 156mg (0.76 mmol) of Dicyclohexylcarbodiimide (DCC), 11mg (0.05 mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) are dissolved in 2mL of dichloromethane at room temperature, and the mixture is reacted at room temperature under nitrogen protection for 12 hours. Insoluble matter was removed by suction filtration, the filtrate was extracted with water, dichloromethane (5 ml×3), the organic phases were combined, washed 1 time with saturated brine (2 mL), dried over anhydrous sodium sulfate, filtered, the solvent was evaporated, sand was produced, and column chromatography was performed to separate and purify (DCM: meoh=60:1 to 40:1) to give a white solid S1187 mg.

¹ H NMR(300MHz,Chloroform-d)δ7.45(dd,J＝6.5,2.1Hz,1H),7.30(dd,J＝6.9,1.8Hz,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 examples 2 to 4 below were obtained by the synthesis method of example 1, and only the corresponding raw materials were replaced.

Example 2: synthesis of Compound S2

¹ H NMR(300MHz,Chloroform-d)δ7.46(dd,J＝6.4,2.1Hz,1H),7.30(dd,J＝6.9,1.8Hz,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

¹ H NMR(300MHz,Chloroform-d)δ7.45(dd,J＝6.5,2.1Hz,1H),7.30(dd,J＝6.8,1.8Hz,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

¹ H NMR(300MHz,Chloroform-d)δ7.45(dd,J＝6.5,2.1Hz,1H),7.31(dd,J＝6.9,1.8Hz,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 (0.50 mmol, synthetic method see example 1) of glycine vincamine, 1008mg (0.50 mmol) of monomethoxy hydroxy polyethylene glycol (MPEG-OH, average molecular weight 2000), 156mg (0.76 mmol) of Dicyclohexylcarbodiimide (DCC), 11mg (0.05 mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) are dissolved in 10mL of dichloromethane at room temperature, protected with nitrogen and reacted at room temperature for 12 hours. Insoluble matters are removed by suction filtration, dichloromethane is removed by rotary evaporation of filtrate, 250mL of diethyl ether is added, stirring is carried out for 1 hour, filtration is carried out, filter cakes are collected, and the crude product is obtained by vacuum drying. The crude product was dialyzed to give S5610 mg as a white solid.

¹ H 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)。

Compounds S6 to S22, S27 to S30 and S39 to S42 in examples 6 to 22, 27 to 30 and 39 to 42 can be obtained by the synthesis method of example 5, and only the corresponding amino acid (hydroxypropionic acid) and monomethoxy hydroxy polyethylene glycol having different molecular weights are required to be replaced.

Example 6: synthesis of Compound S6

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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 (0.50 mmol, synthetic method see example 1) of glycine vincamine, 1008mg (0.50 mmol) of dihydroxypolyethylene glycol (OH-PEG-OH, average molecular weight 2000), 156mg (0.76 mmol) of Dicyclohexylcarbodiimide (DCC), 11mg (0.05 mmol, catalytic amount) of 4-Dimethylaminopyridine (DMAP) were dissolved in 10mL of dichloromethane at room temperature, protected with nitrogen and reacted at room temperature for 12 hours. Insoluble matters are removed by suction filtration, dichloromethane is removed by rotary evaporation of filtrate, 250mL of diethyl ether is added, stirring is carried out for 1 hour, filtration is carried out, filter cakes are collected, and the crude product is obtained by vacuum drying. The crude product was dialyzed to give white solid S23600 mg.

¹ H 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 examples 24 to 26 below were obtained by the synthesis method of example 23, and only the corresponding amino acids and the dihydroxypolyethylene glycols having different molecular weights were used.

Example 24: synthesis of Compound S24

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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 (845 mg,2.85 mmol) was dissolved in anhydrous CH ₂ Cl ₂ (30 mL) was added L-valine tert-butyl hydrochloride (659 mg,1.42 mmol), HATU (2.16 g,5.70 mmol), HOAt (1.38 g,10.15 mmol), DIPEA (1.98 mL,11.4 mmol), and after reaction at room temperature for 7 hours, the reaction progress was checked by TLC plate, and after completion, saturated NH was added ₄ Cl solution is extracted and deactivated, and the organic phase is sequentially treated with saturated NaHCO ₃ Washing with water and saturated salt water, and then using anhydrous Na ₂ SO ₄ Drying, filtering, evaporating the solvent under reduced pressure, and performing column chromatography to obtain Fmoc-protected dipeptide as white solid. The Fmoc-protected dipeptide (1.11 g,2.45 mmol) obtained was dissolved in MeCN (200 mL), piperidine (10 mL) was added at room temperature for 3 hours, the solvent was distilled off under reduced pressure, and the Fmoc-removed dipeptide white solid compound (270 mg, 41%) obtained was purified by column chromatography, and the Fmoc-removed dipeptide obtained was reacted with vincamine demethyl product and monomethoxy-hydroxy polyethylene glycol, respectively, according to the methods of step two and step three in example 1 to obtain compound S31.

¹ H 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)。

Compounds S32 to S38 in examples 32 to 38 below can be obtained by the synthesis method of example 31, and only the corresponding amino acid and the monomethoxy hydroxy polyethylene glycol with different molecular weights are required to be replaced.

Example 32: synthesis of Compound S32

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

Glycine vincamine 440mg (1.10 mmol, synthetic method see example 1), dihydroxypolyethylene glycol (OH-PEG-OH, average molecular weight 2000) 1008mg (0.50 mmol), dicyclohexylcarbodiimide (DCC) 248mg (1.20 mmol), 4-Dimethylaminopyridine (DMAP) 11mg (0.05 mmol, catalytic amount) were dissolved in 10mL dichloromethane at room temperature, nitrogen-protected, and reacted at room temperature for 12 hours. Insoluble matters are removed by suction filtration, dichloromethane is removed by rotary evaporation of filtrate, 250mL of diethyl ether is added, stirring is carried out for 1 hour, filtration is carried out, filter cakes are collected, and the crude product is obtained by vacuum drying. The crude product was dialyzed to give white solid S43730 mg.

¹ H 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)。

Compounds S44 to S56 in examples 44 to 56 below can be obtained by the synthesis method of example 43, and only the corresponding amino acid and the dihydroxypolyethylene glycol having different molecular weights are required to be replaced.

Example 44: synthesis of Compound S44

¹ H 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＝6.8Hz,6H),1.29–1.06(m,4H),0.94(td,J＝1.0,7.2Hz,6H)。

Example 45: synthesis of Compound S45

¹ H 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

¹ H 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

¹ H 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

¹ H 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＝6.8Hz,6H),1.29–1.06(m,4H),0.94(td,J＝1.0,7.2Hz,6H)。

Example 49: synthesis of Compound S49

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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＝6.8Hz,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

¹ H 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＝6.7Hz,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

¹ H 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

¹ H 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

Glycine-alanine linked vincamine 470mg (1.10 mmol, synthetic method see example 31), dihydroxypolyethylene glycol (OH-PEG-OH, average molecular weight 2000) 1008mg (0.50 mmol), dicyclohexylcarbodiimide (DCC) 248mg (1.20 mmol), 4-Dimethylaminopyridine (DMAP) 11mg (0.05 mmol, catalytic amount) were dissolved in 10mL dichloromethane at room temperature, nitrogen protected, and reacted at room temperature for 12 hours. Insoluble matters are removed by suction filtration, dichloromethane is removed by rotary evaporation of filtrate, 250mL of diethyl ether is added, stirring is carried out for 1 hour, filtration is carried out, filter cakes are collected, and the crude product is obtained by vacuum drying. The crude product was dialyzed to give white solid S57740 mg.

¹ H 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＝6.8Hz,6H),1.29–1.07(m,4H),0.94(t,J＝7.3Hz,3H),0.91(t,J＝7.3Hz,3H)。

Compounds S58 to S65 in examples 58 to 65 below can be obtained by the synthesis method of example 57, and only the corresponding amino acid and the dihydroxypolyethylene glycol having different molecular weights are required to be replaced.

Example 58: synthesis of Compound S58

¹ H 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＝6.8Hz,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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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

¹ H 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 velocity and sensory deficit of diabetic peripheral neuropathy model mice

The invention takes PEGylated vincamine series 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 vincamine derivative Vin3 on the motor nerve conduction velocity and pain response of a model mouse with peripheral neuropathy of diabetes on STZ-induced type 1 diabetes mice and type 2 diabetes db/db mice. Experimental results show that Vin3 can obviously improve the motor nerve conduction velocity and the sensory deficit phenomenon of a diabetic peripheral neuropathy model mouse.

1. Principle of experiment

Diabetic peripheral neuropathy involves sensory neurons first, and the main pathological changes are axon atrophy degeneration and even disappearance. It is believed that the therapeutic effect against diabetic peripheral neuropathy can be achieved by protecting and promoting the growth of sensory neuron protrusion. In addition, diabetic peripheral neuropathy patients can have symptoms such as reduced nerve conduction speed, hypoesthesia, hypodynamia and the like. The behavioral indexes such as the motor nerve conduction speed, the mechanical pain threshold value and the thermal pain response time are detected on a diabetic peripheral neuropathy model mouse, and the method can be used for evaluating the improvement effect of the vincamine derivative Vin3 on the individual behavioral level of the diabetic peripheral neuropathy; the growth of the neurites of the peripheral neurons of the mice after the administration is detected, and the protection effect of the compound Vin3 on the neurites of the peripheral neurons can be evaluated.

2. Experimental materials and methods

1) Grouped dosing conditions for animals

Type 1 diabetic mice: male 8 week old C57BL/6J mice were purchased from Experimental animal technologies Inc. of Beijing Vitolihua. After one week of adaptive feeding, the STZ (150 mg/kg) was injected intraperitoneally, and after one week, blood was collected from the tail vein to measure blood glucose, and mice with random blood glucose greater than 16mmol/L were selected. STZ was grouped according to body weight and blood glucose 6 weeks after injection, respectively: control (non-diabetic mice), model (DPN mice), vin3 (40 mg/kg) and continuous gavage for 4 weeks.

Type 2 diabetic mice: male 18 week old BKS db/db mice were purchased from Jiangsu Jiujiaku Biotech Co. After one week of adaptive feeding, the animals were grouped according to body weight and blood glucose, respectively: control group (db/m mice), model group (db/db mice), vin3 administration group (40 mg/kg), and continuous gavage administration for 4 weeks.

2) Motor nerve conduction velocity detection experiment

Before the administration, motor nerve conduction velocity (Motor nerve conduction velocity, MNCV) on both sides of each group of mice was measured at 2 weeks and 4 weeks of administration, and the two-side velocity was averaged as the nerve conduction velocity of each mouse. The specific experimental method comprises the following steps: (1) electrode placement: 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. (2) Calculation of motor nerve conduction velocity: the far end and the near end of the nerve trunk are stimulated in a superstrong mode, 2 times of compound muscle action potentials can be recorded on the innervated muscle, different latencies are measured, and the distance between the far end and the near end is divided by the latency difference between the two points, namely the conduction speed of the nerve. The calculation formula is as follows: nerve conduction velocity (m/s) =distance between two points (cm) ×10/latency difference between two points (ms).

3) Mechanical pain threshold detection experiment

The experiment is based on the principle that rodents' paws retract back when mechanically stimulated. Von frey tactile measurement kit is available from Ugo Basile company, von frey filaments can provide a stimulating force of 0.008g-300g, and the thickness of the filaments determines the amount of stimulating force provided. The specific experimental method comprises the following steps: the fiber yarn with proper thickness is selected to perpendicularly stimulate the skin, and the fiber yarn is replaced to adjust the stimulation force until the fiber yarn is bent. One mouse was assayed 6 times to determine the threshold, according to 50% threshold= (10 x ^f +kδ))/10000 the mechanical pain threshold of 50% was calculated for mice and examined weekly before and after dosing, respectively.

4) Thermal pain latency detection experiment

The response of the animals to infrared thermal stimuli was examined weekly before and after dosing using an infrared plantar pain meter (37370, available from Ugo Basile corporation) to characterize the animals' thermal pain sensitivity. The mice are put into a single cage matched with a machine in advance to adapt to 20min, then the soles of the mice are thermally stimulated, the temperature gradually rises along with the prolongation of time, and when the feet of the mice are lifted or moved away from a heat source, the machine automatically stops and records the thermal stimulation time, namely the thermal pain incubation period. The thermal pain sensitivity was evaluated by comparing the length of the thermal stimulus time.

5) Dorsal root ganglion neuron cell axon growth experiment

After the end of the administration, the fresh dorsal root ganglion (Dorsal root ganglion, DRG) neurons of the mice were taken and digested into single cell suspensions, which were inoculated into culture plates for adherent culture overnight. The beta-tubulin III (Sigma) protein of the DRG neuron primary cells was labeled by immunofluorescence staining. Fluorescence microscopy (Leica) imaging was followed by tracing the total axon length of individual DRG neuronal cells with Neuro J plug-in Image J software.

3. Experimental results

As shown in tables 1-3, vincamine derivative Vin3 was able to significantly improve motor nerve conduction velocity and sensory deficit symptoms in type 1 diabetic peripheral neuropathy mice. All experimental data for this work are shown in mean± SD., and all bar graphs and plots are plotted and analyzed using GraphPad Prism 8 software. T test was used between the two sets of data for significance analysis. ^* 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 dosing group vs. model group.

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

Table 2 mechanical pain threshold 50% (mean± SD., n= 7,g)

TABLE 3 thermal pain latency (mean.+ -. SD., n= 7,s)

The results are shown in tables 4-6, and vincamine derivative Vin3 is capable of significantly improving motor nerve conduction velocity and sensory deficit symptoms in type 2 diabetic mice. T test was used between the two sets of data for significance analysis. ^* 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 treated vs. db/db.

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

Table 5 mechanical pain threshold of 50% (mean± SD., n= 7,g)

TABLE 6 thermal pain latency (mean.+ -. SD., n= 7,s)

As shown in fig. 1, the vincamine derivative Vin3 can significantly improve the axon growth of the DRG neurons of the type 1 diabetic peripheral neuropathy model mouse (fluorescence and quantification). Statistics were tested using T-test, n=5, ^* p<0.05， ^** p<0.01， ^*** p<0.001, respectively compared to the model set. As shown in fig. 2, vincamine derivative Vin3 was also able to significantly promote axon growth of DRG neurons of type 2 diabetic peripheral neuropathy model mice (fluorescence and quantification). Statistics were tested using T-test, n=7, ^* p<0.05， ^** p<0.01， ^*** p<0.001, respectively compared to the model set. The vincamine derivative Vin3 has a protective effect on the axon of peripheral sensory neurons.

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 foot ulcer wound of a diabetic foot model rat on the STZ-induced diabetic rat. Experimental results show that after foot ulcers of diabetic rats are treated by the compound Vin3, the healing of foot ulcer wounds can be obviously promoted, and the injury area is reduced.

1. Principle of experiment

Diabetic foot ulcers (Diabetic foot ulcer, DFU) are a common manifestation of diabetic feet and are also a leading cause of amputation in diabetic patients. The experiment causes a certain area of wound on the instep of a diabetic rat at one time to form a diabetic foot ulcer model, and then the compound Vin3 is given for treatment. The effect of vincamine derivative Vin3 on diabetic foot ulcer wound healing was assessed by photographing the wound and quantifying its area size.

2. Experimental materials and methods

1) Establishment of diabetic rat model

Male 5 week old Sprague Dawley rats were purchased from Experimental animal technologies Inc. of Lerio, beijing. After one week of adaptive feeding, STZ (100 mg/kg) was injected intraperitoneally, and blood glucose was collected from the tail vein after 72 hours, and mice with random blood glucose greater than 16.7mmol/L were selected.

2) Establishment of diabetic foot ulcer rat model

Inhalation anesthesia machine was used after STZ injection for 4 weeks, isoflurane inhalation induced anesthesia, and a complete circular skin wound was created on the back instep of diabetic rats by a disposable 5mm skin biopsy punch and Westcott scissors. After the diabetic foot ulcer rat model was established, it was divided into a control group (non-diabetic rat), a model group (DFU rat) and a Vin3 administration group (40 mg/kg) according to blood sugar and body weight indexes, and was continuously administered by stomach infusion for 4 weeks. 3) Diabetic foot rat ulcer area observation

The size of foot ulcer wound area was observed for each mouse on day 0, day 3, day 7, day 14, day 21 and day 28, respectively, after dosing. The injury area of the foot of each rat was measured with a ruler, and after the Image was taken with a camera, the wound area was quantified by scaling with a scale, image J software.

3. Experimental results

As shown in fig. 3, vincamine derivative Vin3 can obviously promote healing of diabetic foot ulcer wound. Wound area quantification results as shown in table 7, 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 dosing group vs. model group.

Table 7 foot ulcer wound area (mean± SD., n=7, mm ² )

。

Example 3: vincamine derivative Vin3 reduces bleomycin-induced pulmonary fibrosis in mice

The invention utilizes Micro CT to detect the protective effect of the vincamine derivative Vin3 on the pulmonary fibrosis tissue of mice on a bleomycin-induced pulmonary fibrosis model mouse. Experimental results show that administration of the compound Vin3 treatment can significantly reduce the pulmonary fibrosis degree of mice.

1. Principle of experiment

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

2. Experimental materials and methods

Male 7 week old C57BL/6J mice were purchased from Beijing vitamin Torilhua and examined for lung development with micro-CT after one week of adaptive feeding. Mice were divided into a blank group (normal mice), a model group and a Vin3 administration group (100 mg/kg) according to the size of the normal tissue volume of the lung. Then, an inhalation anesthesia machine is adopted, isoflurane inhalation is adopted to induce anesthesia, a model group and an administration group mice are instilled with 5mg/kg of bleomycin solution through an oral trachea, a pulmonary fibrosis model is established, and a control group instills an equal volume of bleomycin solvent. The administration was started the next day, and the administration was continued by gastric lavage 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 pulmonary fibrosis conditions of the mice are observed.

3. Experimental results

As shown in fig. 4, the vincamine derivative Vin3 can obviously reduce the pulmonary fibrosis degree of mice, and has a certain protection effect on pulmonary tissues. After 20 days of administration, the normal tissue volume of the lungs of mice in the administration group was significantly larger than that in the model group. The quantitative results of the normal tissue volumes in the lungs of mice are shown in Table 8, and a T test was used between the two sets of data for significance analysis. ^* 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 dosing 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, the scope of which is defined by the claims. Various modifications and equivalent arrangements of this invention will occur to those skilled in the art, and are intended to be within the spirit and scope of the invention.

Claims

1. A vincamine PEG derivative, which is characterized by comprising a compound shown as a formula I and a formula II; the structural formula of the compound is shown as follows:

in formula I and formula II:

l is

X is

Y is

t is 1 or 2;

R ^1-1 is hydroxyl;

n is an integer of 0 to 100;

r is-O (R) ² )；

R ² Is hydrogen or C _1-6 An alkyl group.

2. The vincamine PEG-derivative of claim 1, wherein the compound of formula i is any one of the following:

。

3. a process for the preparation of a compound of formula i according to claim 1, which comprises the following steps: in a solvent dichloromethane, under the action of alkali dicyclohexylcarbodiimide and 4-dimethylaminopyridine, carrying out condensation reaction between a compound shown in a formula III and a compound shown in a formula IV;

Wherein L and R are as defined in claim 1.

4. A process for the preparation of a compound of formula ii according to claim 1, which comprises the following steps: in a solvent dichloromethane, under the action of alkali dicyclohexylcarbodiimide and 4-dimethylaminopyridine, carrying out condensation reaction between a compound shown in a formula V and a compound shown in a formula III;

wherein X and Y are as defined in claim 1.

5. A pharmaceutical composition comprising a compound of formula i and formula ii as defined in claim 1 or 2, a pharmaceutically acceptable salt thereof.

6. Use of a compound of formula i and formula ii, as claimed in claim 1 or 2, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prophylaxis or treatment of diabetic complications; the diabetes is type 1 diabetes and type 2 diabetes.

7. Use of a compound of formula i or formula ii as defined in claim 1 or 2, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the prophylaxis or treatment of pulmonary fibrosis.