阅读说明：本技术 糖胺聚糖衍生物及其制备方法和用途 (Glycosaminoglycan derivatives, process for their preparation and their use ) 是由 任丽鸽 王景文 金学文 张利利 林森茂 李锂 于 2019-01-29 设计创作，主要内容包括：公开了一种糖胺聚糖羧基化衍生物、其制备方法,及其用于抑制肿瘤生长和/或转移的用途。(Glycosaminoglycan carboxylated derivatives, methods for their preparation, and their use for inhibiting tumor growth and/or metastasis are disclosed.)

A glycosaminoglycan derivative comprising a structural unit of formula (I), a structural unit of formula (IV), and a structural unit of formula (V):

wherein:

R^1m、R¹ⁿand R^1xEach occurrence is independently selected from H, -SO₃ ^-·(1/q E^q+) And- (C ═ O) CH₃And R is^1m、R¹ⁿAnd R^1xIn each occurrence is preferably-SO₃ ^-·(1/q E^q+) Or- (C ═ O) CH₃；

R^2m、R²ⁿAnd R^2xEach independently at each occurrence is selected from H and-SO₃ ^-·(1/q E^q+)；

R³ⁿEach independently at each occurrence is selected from H and-SO₃ ^-·(1/q E^q+)；

R^4m、R⁴ⁿAnd R^4xEach independently at each occurrence is selected from H and-SO₃ ^-·(1/q E^q+)；

E is independently selected at each occurrence from H, an alkali metal (preferably lithium, sodium, potassium, rubidium or cesium), an alkaline earth metal (preferably magnesium or calcium), and aluminum;

q is, independently for each occurrence, an integer of 1, 2 or 3;

the weight average molecular weight of the glycosaminoglycan derivative is 7000-14000Da, preferably 8000-13500Da, such as 8500-13000Da, 8500-12500Da or 9000-12500 Da; and is

The glycosaminoglycan derivatives have an uronic acid ring opening degree of 25-80%, preferably 25-60%.

The glycosaminoglycan derivative of claim 1 further comprising structural units of the formula (II):

wherein:

R^2tand R^3tEach independently at each occurrence is selected from H and-SO₃ ^-·(1/q E^q+)；

E is independently selected at each occurrence from H, an alkali metal (preferably lithium, sodium, potassium, rubidium or cesium), an alkaline earth metal (preferably magnesium or calcium), and aluminum; and is

q is, independently for each occurrence, an integer of 1, 2 or 3.

The glycosaminoglycan derivative of claim 1 or 2 further comprising structural units of the formula (III):

wherein:

R^2peach independently at each occurrence is selected from H and-SO₃ ^-·(1/q E^q+)；

E is independently selected at each occurrence from H, an alkali metal (preferably lithium, sodium, potassium, rubidium or cesium), an alkaline earth metal (preferably magnesium or calcium), and aluminum; and is

q is, independently for each occurrence, an integer of 1, 2 or 3.

The glycosaminoglycan derivative of any one of claims 1 to 3 further comprising structural units of the formula (VI):

wherein:

R^2rand R^3rEach independently at each occurrence is selected from H and-SO₃ ^-·(1/q E^q+)；

E is independently selected at each occurrence from H, an alkali metal (preferably lithium, sodium, potassium, rubidium or cesium), an alkaline earth metal (preferably magnesium or calcium), and aluminum; and is

q is, independently for each occurrence, an integer of 1, 2 or 3.

The glycosaminoglycan derivative of any one of claims 1 to 4 further comprising structural units of the formula (VII):

wherein:

R^1seach occurrence is independently selected from H, -SO₃ ^-·(1/q E^q+) And- (C ═ O) CH₃And is preferably-SO₃ ^-·(1/q E^q+) Or- (C ═ O) CH₃；

R^2s、R^3sAnd R^4sEach independently at each occurrence is selected from H and-SO₃ ^-·(1/q E^q+)；

E is independently selected at each occurrence from H, an alkali metal (preferably lithium, sodium, potassium, rubidium or cesium), an alkaline earth metal (preferably magnesium or calcium), and aluminum; and is

q is, independently for each occurrence, an integer of 1, 2 or 3.

Glycosaminoglycan derivative according to any of the claims 1 to 5, wherein the uronic acid epoxy degree is less than 25%.

The glycosaminoglycan derivative of any one of claims 1 to 6 wherein the glycosaminoglycan derivative has a molecular weight distribution as follows:

molecular weight Range (Da) Proportion (wt%) Greater than 10000 15-80, preferably 25-80 6000-10000 15-50 Less than 6000 5-50

Preferably, the molecular weight distribution of the glycosaminoglycan derivative is as follows:

molecular weight Range (Da) Proportion (wt%) Greater than 10000 30-75 6000-10000 20-40 Less than 6000 5-30

Or

Molecular weight Range (Da) Proportion (wt%) Greater than 10000 30-75 6000-10000 20-40 Less than 6000 0-30

。

The glycosaminoglycan derivative of any one of claims 1 to 7 wherein the glycosaminoglycan derivative has a ratio of sulfonic to carboxylic acids of 0.80 to 1.65, preferably 1.0 to 1.4.

A pharmaceutical composition comprising a prophylactically or therapeutically effective amount of the glycosaminoglycan derivative of any one of claims 1 to 8 and a pharmaceutically acceptable carrier, preferably the pharmaceutical composition is a solid formulation, a semi-solid formulation, a liquid formulation, or a gaseous formulation.

Use of a glycosaminoglycan derivative according to any one of claims 1 to 8 for the preparation of a medicament for the inhibition of tumor growth and/or metastasis.

The use of claim 10, wherein the tumor is a solid tumor, a hematologic tumor, or a soft tissue tumor; and preferably a solid tumor such as breast, pancreatic, bladder, prostate, colon, gastric or lung cancer.

A method for preparing the glycosaminoglycan derivative of any one of claims 1 to 8, comprising the steps of:

a) optionally, C2, C3 epoxidation of the uronic acid residue of the glycosaminoglycan, which is preferably performed in aqueous alkaline solution (preferably aqueous sodium hydroxide solution);

b) optionally, hydrolytic opening of the epoxidation product obtained in step a), preferably under neutral conditions;

c) in the presence of a vicinal diol and optionally a vicinal OH/NH₂Oxidizing 10% to 100% (preferably 25% to 100%) of the 2-O-and optionally the 2N-, 3-O-non-sulfated residues of the glycosaminoglycan, preferably by periodate (preferably sodium periodate), under conditions effective to convert the glycosaminoglycan to a dialdehyde; and

d) further oxidation of the product from step c), preferably by means of a chlorite salt (preferably sodium chlorite), under conditions effective to convert the dialdehyde to carboxyl groups and without nitrogen protection;

preferably, the method further comprises a step of 2N-desulfation of glucosamine residues, which is carried out before step a), after step a) or after step b), said desulfation step comprising salt formation with pyridine followed by stirring in a mixed solvent of DMSO and water or methanol.

The method according to claim 12, wherein said glycosaminoglycan is a natural heparin or a synthetic heparin (optionally chemically or enzymatically modified) from any animal and organ source, preferably selected from the group consisting of optionally 2-O-and/or 2-N-desulphated heparin, unfractionated heparin, Low Molecular Weight Heparin (LMWH) and sulfated heparin, having a sulfonic to carboxylic ratio of 0.8-2.8, preferably 0.9-2.5; more preferably selected from unfractionated heparin and LMWH, optionally 2-O-and/or 2-N-desulphated.

The method of claim 12 or 13, wherein the glycosaminoglycan has a weight average molecular weight of 10000Da to 30000Da, preferably 15000Da to 25000Da, such as 15000Da to 20000Da, 15000Da to 19000Da or 17000Da to 19000 Da.

The method of any one of claims 12 to 14, wherein the glycosaminoglycan derivative exhibits a carboxyl group increment of 1.3 to 2.0.