WO2023227069A1 - Polidocanol with single molecular weight, and use thereof - Google Patents

Abstract

The present invention relates to use of polidocanol with a single molecular weight in the preparation of a drug for treating disease. The polidocanol can serve as a curing agent and/or a hemostatic agent for treatment. The polidocanol with a single molecular weight is prepared by the following preparation method: reacting polyethylene glycol with a single molecular weight with 1-halododecane, and performing separation and purification to obtain the polidocanol with a single molecular weight. The propolidocanol of the present invention is produced by a stable process, is of controllable quality, is good in efficacy, and causes few toxic and side effects.

Description

A single molecular weight polydocanol and its application Technical field

The invention belongs to the technical field of pharmaceutical preparations, and specifically relates to a preparation method of single molecular weight polidocanol and its application in preparing disease treatment drugs.

Background technique

Polidocanol, also known as ethoxyhardened alcohol and lauromacrol, is a polymer product obtained by polymerizing lauryl alcohol and multiple ethylene oxides. At present, polidocanol is mainly produced through the polymerization reaction of lauryl alcohol and ethylene oxide under alkali-catalyzed conditions. The quality and stability of polidocanol products are greatly affected by the purity of starting materials, catalysts, reaction pressure, temperature and post-treatment processes. On the one hand, commercially available lauryl alcohol contains many impurities. In addition to decanol and tetradecanol, there are also many unknown impurities. The by-products generated after these impurities are introduced into the polymerization reaction of polidocanol are difficult to separate from polidocanol, which will directly affect the quality of the finished product of polidocanol. On the other hand, in addition to unreacted lauryl alcohol and ethylene oxide, many by-products will be produced during the synthesis of polydocanol. The main by-products are free polyethylene glycol, diethylene glycol, and dioxane. , formaldehyde, acetaldehyde, ethylene glycol, etc. These impurities have significant adverse effects on the safety and effectiveness of polidocanol.

The patent with the publication number CN103922901 discloses a purification method of polidocanol. The high-purity polidocanol refined product is finally obtained through vacuum distillation before adjusting the pH value of the crude product. However, this invention cannot avoid the risk of by-products generated by impurities (such as n-decanol and n-tetradecanol) brought into lauryl alcohol and ethylene oxide.

The patent with publication number CN100335453C discloses a preparation method of polidocanol, which uses NaOH as the catalyst, and adjusts the pH of the crude product to 5-7 with acetic acid to obtain a white or off-white ointment finished product. The finished product prepared by the present invention cannot fully meet pharmaceutical grade quality standards.

The patent with publication number CN113200821A discloses a purification method of lauryl alcohol and a synthesis method of polidocanol. High-purity lauryl alcohol is prepared by vacuum distillation, but the distillation purification requirement is -0.08MPa, and the distillation temperature 130～150℃. In the process of preparing polidocanol, the dehydration temperature of lauryl alcohol is 90-125°C, the vacuum degree is -0.08MPa, the polymerization temperature is 100-115°C, and the polymerization pressure is 0.1-0.8MPa. After the polymerization is completed, the degassing temperature is 90-0.08MPa. 120℃. It can be seen that the polymerization reaction is carried out under high temperature and negative pressure conditions, which has high requirements on equipment and technology.

The patent with publication number CN113527060A discloses a refining process of lauryl alcohol and a process of preparing lauromanol using the refined product as raw material. This patent uses an organic solvent to refine the raw material of lauryl alcohol through recrystallization. After dissolving the organic solvent and lauryl alcohol, it needs to be cooled to -5℃~10℃, stirred and crystallized, recrystallized, filtered and then distilled under reduced pressure (-0.1MPa~0.05 MPA), temperature 70~100℃. In the preparation process of lauronanol, the polymerization temperature of lauryl alcohol and ethylene oxide is 130~180℃, and the pressure is 0.1~0.5MPa. It can be seen that the preparation of lauryl alcohol and lauronanol has strict requirements on temperature and pressure, as well as high requirements on equipment and technology.

Polidocanol is a surfactant-type hardener whose structure is very similar to the cell membrane phospholipid bilayer. By interfering with the surfactant substances on the cell membrane, it destroys the cell membrane structure, causing the lysis and death of vascular endothelial cells, and then hair Vascular fibrosis and vascular occlusion. It has fixed hydrophilic ends and hydrophobic ends at the same time, which can be oriented and arranged on the surface of the solution, significantly reducing the surface tension of the solution and forming foam. Therefore, polidocanol is widely used in foam sclerosis treatment. At present, polidocanol is mainly used in sclerotherapy for varicose veins, venous malformations, hemangiomas, internal hemorrhoids, cystic diseases, acne, tumor ablation, etc.

However, since the existing polidocanol is a mixture of different molecular weights, polidocanols with different chain lengths have different foaming effects and medicinal effects, making it difficult to control during the application process. Therefore, it is of great significance to develop and prepare polidocanol with a single molecular weight to effectively control and improve the quality of polidocanol to improve the efficacy of polidocanol.

Contents of the invention

In order to solve the above problems, the technical solution adopted by the present invention is:

The present invention provides the application of single molecular weight polidocanol in the preparation of disease treatment drugs, and the polidocanol is used as a sclerosing agent and/or hemostatic agent for treatment.

Further, the diseases include cystic diseases, vascular related diseases, tumors, obstetric and gynecological diseases, inflammation and/or pain;

Preferably, the vascular related diseases include varicose veins, vascular malformations and/or hemorrhage;

Preferably, the obstetric and gynecological diseases include uterine submucosal fibroids and cesarean scar pregnancy;

Preferably, the cystic disease includes abdominal organ cysts, body surface cysts, lymphoceles, thyroid cysts, bone cysts, digital mucocele and/or pelvic encapsulated effusion;

Preferably, the tumors include benign or malignant solid tumors;

Preferably, the inflammation includes infectious inflammation and/or non-infectious inflammation;

Preferably, the pain includes chronic pain, acute pain and/or cancer pain.

Further, the tumors include: infantile hemangioma, congenital hemangioma, plexiform hemangioma, spindle cell hemangioma, epithelioid hemangioma, pyogenic granuloma, Kaposiform hemangioendothelioma, reticular hemangioma Endothelioma, papillary intralymphatic hemangioendothelioma, composite hemangioendothelioma, Kaposi's sarcoma, angiosarcoma, epithelioid hemangioendothelioma, uterine fibroids and/or abdominal visceral malignancies;

Preferably, the pain includes: muscle and soft tissue pain, bone and joint pain and/or visceral pain;

Preferably, the inflammation includes: tendinitis, tendinitis, tendinosis, peritendinitis, ankylosing spondylitis, sacroiliitis, gouty arthritis, myofasciitis, tenosynovitis and/or frozen shoulder;

Preferably, the varicose veins include: varicose veins of lower limbs, esophageal and gastric varices, varicocele, abdominal wall varicose veins and/or hemorrhoids;

Preferably, the bleeding includes: gastrointestinal bleeding, bladder bleeding, bleeding caused by early, middle or late cancer;

Preferably, the abdominal organ cysts include: liver cysts, renal cysts, parapelvic cysts, pancreatic cysts, endometrium Ectopic cysts and/or ovarian chocolate cysts;

Preferably, the body surface cysts include: skin cysts and/or finger mucous cysts;

Preferably, the abdominal organ malignant tumors include liver cancer.

Further, the molecular structure of the single molecular weight polydocanol monomer is:

Where n is an integer, 4≤n≤24;

Preferably, n is an integer, 8≤n≤15;

Preferably, n is selected from any of 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 A numerical value, each value of n corresponds to a polidocanol product of a defined molecular weight.

More preferably, n is 8, 9, 10 or 11.

Further, the single molecular weight polydocanol is prepared by the following preparation method: reacting single chain length polyethylene glycol with 1-halododecane, separating and purifying the product to obtain single molecular weight polydocanol. alcohol.

In the present invention, "single molecular weight" means that the compound of the present invention is not a mixture of homologues with different molecular weights. It refers to the average molecular weight of a polymer in which the molecular weight of the compound of the present invention is not a mixture of homologues with different molecular weights.

For example, when the polydocanol with a single chain length of the present invention contains PEG4 (four ethylene glycol units), the polydocanol with a single chain length only contains PEG4, excluding those containing PEG3, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, Polydocanol of other PEGs such as PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51;

When the polydocanol with a single chain length of the present invention contains PEG5 (five ethylene glycol units), the polydocanol with a single chain length only contains PEG5, excluding those containing PEG3, PEG4, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, Polydocanol of other PEGs such as PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG6 (six ethylene glycol units), the polydocanol with a single chain length Alcohol only contains PEG6, excluding alcohol such as PEG3, PEG4, PEG5, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, Polidocanol from other PEGs such as PEG49, PEG50 and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG7 (seven ethylene glycol units), the polydocanol with a single chain length only contains PEG7, excluding those containing PEG3, PEG4, PEG5, PEG6, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, Polydocanol of other PEGs such as PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG8 (eight ethylene glycol units), the polydocanol with a single chain length only contains PEG8, excluding those containing PEG3, PEG4, PEG5, PEG6, PEG7, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, Polydocanol of other PEGs such as PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG9 (nine ethylene glycol units), the polydocanol with a single chain length only contains PEG9, excluding those containing PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, Polydocanol of other PEGs such as PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG10 (ten ethylene glycol units), the polydocanol with a single chain length only contains PEG10, excluding those containing PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, Polydocanol of other PEGs such as PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG11 (eleven ethylene glycol units), the polydocanol with a single chain length Docanol only contains PEG11, excluding PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, Polidocanol from other PEGs such as PEG48, PEG49, PEG50 and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG12 (twelve ethylene glycol units), the polydocanol with a single chain length only contains PEG12, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG13 (thirteen ethylene glycol units), the polydocanol with a single chain length only contains PEG13, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG14 (fourteen ethylene glycol units), the polydocanol with a single chain length only contains PEG14, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG15 (fifteen ethylene glycol units), the polydocanol with a single chain length only contains PEG15, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , Polydocanol of other PEGs such as PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG16 (sixteen ethylene glycol units), the polydocanol with a single chain length only contains PEG16, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG17 (seventeen ethylene glycol units), the polydocanol with a single chain length only contains PEG17, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG18 (eighteen ethylene glycol units), the polydocanol with a single chain length only contains PEG18, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG19, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG19 (nineteen ethylene glycol units), the polydocanol with a single chain length only contains PEG19, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG20, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG20 (twenty ethylene glycol units), the polydocanol with a single chain length only contains PEG20, excluding those containing PEG3, PEG4, PEG5, and PEG6. , PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG21, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32 , PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50 and PEG51 and other PEG polidocanols.

When the polydocanol with a single chain length of the present invention contains PEG21 (twenty-one ethylene glycol units), the polydocanol with a single chain length only contains PEG21, excluding PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG22, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, Polydocanol of other PEGs such as PEG32, PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG22 (twenty-two ethylene glycol units), the polydocanol with a single chain length only contains PEG22, excluding those containing PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG23, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, Polydocanol of other PEGs such as PEG32, PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG23 (twenty-three ethylene glycol units), the polydocanol with a single chain length only contains PEG23, excluding those containing PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG24, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, Polydocanol of other PEGs such as PEG32, PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, PEG47, PEG48, PEG49, PEG50, and PEG51.

When the polydocanol with a single chain length of the present invention contains PEG24 (twenty-four ethylene glycol units), the polydocanol with a single chain length only contains PEG24, excluding those containing PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, PEG11, PEG12, PEG13, PEG14, PEG15, PEG16, PEG17, PEG18, PEG19, PEG20, PEG21, PEG22, PEG23, PEG25, PEG26, PEG27, PEG28, PEG29, PEG30, PEG31, PEG32, PEG33, PEG34, PEG35, PEG36, PEG37, PEG38, PEG39, PEG40, PEG41, PEG42, PEG43, PEG44, PEG45, PEG46, Polidocanol from other PEGs such as PEG47, PEG48, PEG49, PEG50 and PEG51.

Further, the preparation method of the present invention includes the following steps:

S1: Mix single molecular weight polyethylene glycol with catalyst and organic solvent, add 1-halododecane dropwise, raise the temperature to the preset temperature, and keep the reaction warm;

S2: After the reaction is completed, cool down the reactant. After it reaches room temperature, add water to dilute, extract, and separate the liquids;

S3: Filter, concentrate, dry, separate and purify the organic phase to obtain the single molecular weight polydocanol.

Further, the catalyst is one or more of sodium hydroxide, sodium hydride, potassium hydroxide, potassium hydride, potassium tert-butoxide or sodium tert-butoxide, preferably sodium hydroxide or sodium hydride.

Further, the organic solvent is selected from: ethanol, methanol, isopropyl alcohol, tetrahydrofuran (THF), acetone, acetonitrile, toluene, xylene, n-hexane, cyclohexane, isooctane, n-pentane, petroleum ether, At least one of dimethyl sulfoxide (DSMO), N,N-dimethylformamide, dichloromethane (DCM), and diethoxymethane (DEM). More preferably, it is at least one selected from tetrahydrofuran (THF), dimethyl sulfoxide (DSMO), acetonitrile, toluene, and N,N-dimethylformamide.

Further, the 1-halododecane is 1-chlorododecane, 1-bromododecane or 1-iodododecane, preferably 1-bromododecane.

Further, the preset temperature is 80°C to 95°C, such as 80°C, 81°C, 82°C, 83°C, 84°C, 85°C, 86°C, 87°C, 88°C, 89°C, 90°C, 91°C. , 92°C, 93°C, 94°C, 95°C, preferably 80 to 90°C.

Further, the time of the insulation reaction is 12 to 24 hours, preferably 16 to 24 hours, for example, the time of the insulation reaction is 16, 17, 18, 19, 20, 21, 22, 23 or 24 hours;

More preferably, the insulation reaction time is 16 to 20 hours.

Further, the molar ratio of the single molecular weight polyethylene glycol to the 1-halododecane is 2 to 15:1, more preferably 2 to 10:1, such as 2:1, 3: 1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1.

Further, in the step S2, the extraction operation is repeated 2 to 6 times, more preferably 3 to 5 times, more preferably 3 or 4 times;

Preferably, the extraction solvent is one or a combination of one or more of diethyl ether, ethyl acetate, dichloromethane, chloroform, methyl tert-butyl ether, and toluene. More preferably, the extraction solvent is Ethyl acetate;

Preferably, the organic phase is a mixture of organic phases obtained by the above repeated extraction;

Preferably, in the step S3, the desiccant used for drying is one or more of Na ₂ SO ₄ or MgSO ₄ .

Further, in the step S3, the separation and purification is carried out by chromatography, preferably column chromatography, and the column The eluent used in chromatography is selected from one or more of ethyl acetate, methanol, ethanol, n-hexane, petroleum ether or dichloromethane.

The present invention also provides a preparation of polidocanol, including the aforementioned single molecular weight polidocanol or a pharmaceutically acceptable salt thereof and pharmaceutically acceptable excipients;

Preferably, the pharmaceutically acceptable salt is obtained by adding a pharmaceutically acceptable acid, and the pharmaceutically acceptable acid includes hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid or lactic acid;

Preferably, the preparation also includes active substances and clean gas;

More preferably, the preparation is an injection. Particularly preferably, the injection is a stock solution or a foaming preparation. In the embodiment of the present invention, the injection is a foaming preparation.

The invention also provides a polidocanol foam preparation for use in the preparation and treatment of cystic diseases, varicose veins, vascular malformations, gastrointestinal bleeding, cesarean scar pregnancy, and benign or malignant solid tumors.

The present invention also provides the use of single molecular weight polidocanol in the preparation of disease treatment drugs, and the polidocanol is used as a sclerosing agent and/or hemostatic agent for treatment.

The diseases include cystic diseases, vascular related diseases, tumors, obstetric and gynecological diseases, inflammation and/or pain.

Preferably, the vascular related diseases include varicose veins, vascular malformations and/or hemorrhage.

Preferably, the obstetric and gynecological diseases include uterine submucosal fibroids and cesarean scar pregnancy.

Preferably, the cystic disease includes abdominal organ cysts, body surface cysts, lymphoceles, thyroid cysts, bone cysts, digital mucocele and/or pelvic encapsulated effusion.

Preferably, the tumors include benign or malignant solid tumors.

Preferably, the inflammation includes infectious inflammation and/or non-infectious inflammation.

Preferably, the pain includes chronic pain, acute pain, and cancer pain.

The tumors include: infantile hemangioma, congenital hemangioma, plexiform hemangioma, spindle cell hemangioma, epithelioid hemangioma, pyogenic granuloma, Kaposiform hemangioendothelioma, reticular hemangioendothelioma, Papillary intralymphatic hemangioendothelioma, combined hemangioendothelioma, Kaposi's sarcoma, angiosarcoma, epithelioid hemangioendothelioma, uterine fibroids, and/or abdominal visceral malignancies.

The pain includes muscle and soft tissue pain, bone and joint pain, and/or visceral pain.

The inflammation includes tendonitis, tendonitis, tendinosis, peritendinitis, spondylitis, sacroiliitis, gouty arthritis, myofasciitis, tenosynovitis and/or frozen shoulder.

The tumors include: infantile hemangioma, congenital hemangioma, plexiform hemangioma, spindle cell hemangioma, epithelioid hemangioma, pyogenic granuloma, Kaposiform hemangioendothelioma, reticular hemangioendothelioma, Papillary intralymphatic hemangioendothelioma, combined hemangioendothelioma, Kaposi's sarcoma, angiosarcoma, epithelioid hemangioendothelioma, uterine fibroids, abdominal viscera malignant tumors.

The varicose veins include: varicose veins of the lower limbs, esophageal and gastric varices, varicocele, abdominal wall varicose veins and/or hemorrhoids.

The bleeding includes: gastrointestinal bleeding, bladder bleeding, and bleeding caused by early, middle or late cancer.

The abdominal organ cysts include: liver cysts, renal cysts, pararenal pelvic cysts, pancreatic cysts, endometriosis cysts and/or ovarian chocolate cysts.

The body surface cysts include: skin cysts and finger mucous cysts.

The abdominal organ malignant tumors include liver cancer.

Beneficial effects of the present invention:

The present invention uses single molecular weight polidocanol to prepare drugs for cystic diseases, varicose veins, vascular malformations, gastrointestinal bleeding, cesarean scar pregnancy, and benign or malignant solid tumors. Compared with the prior art, this invention has The invented single molecular weight polidocanol has better efficacy and fewer toxic and side effects.

The preparation process of polidocanol of the present invention overcomes the defects of the existing process that the raw material of lauryl alcohol has many impurities, many side reaction products, and high process conditions (such as pressure, temperature) and equipment requirements. The process is stable, the quality is controllable, and it is suitable for Industrial production.

The single molecular weight polydocanol and its foaming preparation of the present invention eliminate the polydocanol with inappropriate PEG chain length in the original drug, thereby obtaining the single molecular weight polydocanol with better foaming effect than the original drug. Alcohol, foaming effect and foam maintenance time are superior to the foaming preparations of polidocanol in the prior art.

Description of the drawings

Figure 1 shows the hydrogen nuclear magnetic spectrum of DD-PEG4-OH;

Figure 2 shows the HPLC chromatogram of DD-PEG4-OH;

Figure 3 shows the chromatogram of 1% reference substance of DD-PEG4-OH;

Figure 4 shows the mass spectrum of DD-PEG4-OH;

Figure 5 shows the hydrogen nuclear magnetic spectrum of DD-PEG8-OH;

Figure 6 shows the HPLC chromatogram of DD-PEG8-OH;

Figure 7 shows the chromatogram of 1% reference substance of DD-PEG8-OH;

Figure 8 shows the mass spectrum of DD-PEG8-OH;

Figure 9 shows the hydrogen nuclear magnetic spectrum of DD-PEG9-OH;

Figure 10 shows the HPLC chromatogram of DD-PEG9-OH;

Figure 11 shows the chromatogram of 1% reference substance of DD-PEG9-OH;

Figure 12 shows the mass spectrum of DD-PEG9-OH;

Figure 13 shows the TIC chart of DD-PEG9-OH;

Figure 14 shows the hydrogen nuclear magnetic spectrum of DD-PEG10-OH;

Figure 15 shows the HPLC chromatogram of DD-PEG10-OH;

Figure 16 shows the chromatogram of 1% reference substance of DD-PEG10-OH;

Figure 17 shows the mass spectrum of DD-PEG10-OH;

Figure 18 shows the hydrogen nuclear magnetic spectrum of DD-PEG11-OH;

Figure 19 shows the HPLC chromatogram of DD-PEG11-OH;

Figure 20 shows the chromatogram of 1% reference substance of DD-PEG11-OH;

Figure 21 shows the mass spectrum of DD-PEG11-OH;

Figure 22 shows the hydrogen nuclear magnetic spectrum of DD-PEG12-OH;

Figure 23 shows the HPLC chromatogram of DD-PEG12-OH;

Figure 24 shows the chromatogram of 1% reference substance of DD-PEG12-OH;

Figure 25 shows the TIC chart of DD-PEG12-OH;

Figure 26 shows the mass spectrum of DD-PEG12-OH;

Figure 27 shows the hydrogen nuclear magnetic spectrum of DD-PEG24-OH;

Figure 28 shows the HPLC chromatogram of DD-PEG24-OH;

Figure 29 shows the chromatogram of 1% reference substance of DD-PEG24-OH;

Figure 30 shows the mass spectrum of DD-PEG24-OH;

Figure 31 shows the foam emptying time (s) of the sample of Example 3;

Figure 32 shows the foam half-life (s) of the sample of Example 3;

Figure 33 shows the foam coalescence time (m) of the sample of Example 3;

Figure 34 shows images of vein appearance 14 days after injection of different types of polidocanol into the ear margin veins of rabbits in Example 4;

Figure 35 shows the 14-day venous sclerosis effective rate after injecting different types of polidocanol into the ear margin veins of rabbits in Example 4;

Figure 36 shows images of vein appearance 21 days after injection of different types of polidocanol into rabbit ear margin veins in Example 4;

Figure 37 shows the 21-day venous sclerosis effective rate after injecting different types of polidocanol into the ear margin veins of rabbits in Example 4;

Figure 38 shows the pathological changes of the marginal ear vein of rabbits after 14 days of administration in Example 4.

Detailed ways

Unless otherwise defined, all scientific and technical terms used in the present invention have the same meaning as commonly understood by a person skilled in the art to which this invention relates.

In the present invention, the term "sclerotherapy" refers to the injection of sclerosant to turn blood vessels into fibrous cords, which are eventually absorbed by surrounding tissues, or to use sclerosis to destroy cells in the lesions, causing them to dehydrate, coagulate, and denature, thereby producing sterility. Inflammation promotes cyst wall collapse, adhesion, and fibrosis until closure, thereby reducing or eliminating corresponding clinical symptoms.

In the present invention, the term "loading" refers to the operation of adding the product to be separated onto the chromatography column.

In the present invention, "forward separation" refers to the use of column chromatography to separate products in the present invention, in which the stationary phase is polar and the mobile phase is a solvent with less polarity than the stationary phase.

In the present invention, the term "retention time" refers to the time elapsed by the separated sample component from the beginning of the injection to the time when the maximum concentration value of the component appears, that is, from the beginning of the injection to the time when the apex of the chromatographic peak of a certain component appears. The time is called the retention time of this component, expressed as RT, and is often measured in minutes (min) as the time unit.

In the present invention, the term "Tessari method", also known as vortex technology, refers to the production of foam sheets using two disposable plastic syringes, one containing a liquid hardener solution and the other containing air. The port is connected to a three-way valve, and the contents of the two syringes are quickly pushed back and forth multiple times, creating foam through the turbulence created.

In the present invention, the term "HO-PEG24-OH" refers to tetraethylene glycol.

In the present invention, the term "DD-PEG4-OH" refers to polidocanol containing 4 PEGs, "DD-PEG8-OH", "DD-PEG9-OH", "DD-PEG10-OH", "DD -PEG11-OH”, “DD-PEG12-OH”, and “DD-PEG24-OH” are the same.

In the present invention, the terms "10V" and "8V" refer to THF (170mL, 8V). For example, a certain solid material is 1g, and 10V solvent refers to 10ml of solvent. In the embodiment of the present invention, the raw material tetraethylene glycol is directly used as the solvent. However, PEG with other molecular weights, such as PEG8 or above, must be added with a solvent because it is a solid. For example, THF (170 mL, 8V) in the embodiment of the present invention refers to the mass volume ratio of THF to solute. The mass of HO-PEG24-OH Volume ratio.

In the present invention, the term "eq" means "Equivlent", which means molar equivalent.

In the present invention, the term "TLC" refers to thin layer chromatography (Thin Layer Chromatography, TLC), which usually refers to a liquid chromatography method using an adsorbent as a stationary phase. That is, the stationary phase is evenly spread into a thin layer on a smooth surface such as glass, metal, or plastic, and the sample is placed at one end of the thin layer. The mobile phase flows through the stationary phase by capillary action, causing the separated substances to unfold.

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. The described embodiments are only some of the embodiments of the present invention, but not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of protection of the present invention. If the specific conditions are not specified in the examples, the conditions should be carried out according to the conventional conditions or the conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not indicated, they are all conventional products that can be purchased commercially.

Example 1 Synthesis of Single Molecular Weight Polydocanols with Different Chain Lengths

1. Synthesis, separation, purification and characterization of DD-PEG4-OH

NaOH (2g, 50mmol) and tetrabutylammonium bromide (810mg, 50mmol) were added to tetraethylene glycol (97g, 10v), stirred, and 1-bromododecane (12.5g, 50mmol) was dropped at 60°C. Add it to the system, and after the dripping is complete, the system is heated to 80°C and reacted for 12 to 16 hours. The reaction was brought to room temperature, diluted with water (400 ml), extracted three times with ethyl acetate (200 ml), separated, combined organic phases, dried over anhydrous Na ₂ SO ₄ , filtered, concentrated to dryness, column chromatography (elution Liquid ethyl acetate:n-hexane=1:1), 12g of light yellow oily liquid was eluted, and the yield was 66.3%.

The NMR results are shown in Figure 1. ¹ HNMR (300MHz, CDCl ₃ ) δ: 4.536-4.572 (1H, t), 3.314~3.518 (18H, m), 1.454~1.497 (1H, t), 1.249 (18H, s ), 0.839~0.884(3H, t);

Using the main component self-control method, the product content was measured to be 98.99%. The specific results are shown in Table 1. The chromatographic results are shown in Figure 2. Figure 3 is the chromatogram of the 1% reference substance of DD-PEG4-OH. The mass spectrum results are as follows As shown in Figure 4, MS (ESI+): m/z+H=363.4.

Table 1 Chromatogram results of DD-PEG4-OH

2. Synthesis, isolation, purification and characterization of DD-PEG8-OH

Add sodium hydride (2.2g, 55mmol) to a 500ml single-neck bottle containing DMSO (200ml) at room temperature, stir for 30mins, then slowly add PEG8 (20g, 54.1mmol) under nitrogen protection, and stir at room temperature for 2hrs. Under nitrogen protection, 1-bromododecane (13.5g, 54.1mmol) was slowly added into the reaction bottle, and then the reaction was raised to 90°C and stirred for 18hrs. The reaction was brought to room temperature, water (400 ml) was added, and extracted with ethyl acetate (100 ml) for a total of 3 times. The organic phases were combined and washed with saturated sodium chloride solution (100 ml). The organic phase was dried with anhydrous Na ₂ SO ₄ , filter, concentrate to dryness, dissolve dichloromethane and load the sample, forward separation, separation gradient dichloromethane: methanol = 0% ~ 3%, determine the product point with TLC spot plate, combine the product fractions and concentrate to obtain 2.1g of light yellow solid. Yield 7.2%.

The NMR results are shown in Figure 5. ¹ HNMR (300MHz, CDCl ₃ ) δ: 0.87～0.90 (3H, t), 1.25 (18H, s), 1.52～1.59 (2H, t), 2.68 (1H, s), 3.42～3.46(2H,t), 3.55～3.72(29H,m).

Using the main component self-control method, the product content was measured to be 98.62%. The specific results are shown in Table 1. The chromatographic results are shown in Figure 6. Figure 7 is the chromatogram of the 1% reference substance of DD-PEG8-OH. The mass spectrum results are as follows As shown in Figure 8, MS (ESI+): m/z+H=539.5.

Table 2 Chromatogram results of DD-PEG8-OH

3. Synthesis, isolation, purification and characterization of DD-PEG9-OH

Add sodium hydride (484 mg, 12.1 mmol) to a 500 ml single-neck bottle containing dimethyl sulfoxide (200 ml) at room temperature, stir for 30 mins, then slowly add nonaethylene glycol (20 g, 48.3 mmol) under nitrogen protection, and stir at room temperature for 2 hrs. . Under nitrogen protection, 1-bromododecane (3.02g, 12.1mmol) was slowly added into the reaction bottle, and then the reaction was raised to 90°C and stirred for 18hrs. The reaction was brought to room temperature, water (400ml) was added, and extracted with ethyl acetate (100ml) for a total of 3 times. The organic phases were combined and washed with saturated sodium chloride solution (100ml). The organic phase was dried with anhydrous Na2SO4 and filtered. , concentrated to dryness, DCM was dissolved and loaded, forward separation, separation gradient dichloromethane: methanol = 0% ~ 3%, TLC spot plate to determine the product point, product fractions were combined and concentrated to obtain 1.36g of light yellow solid, yield 19.3 %.

The NMR results are shown in Figure 9. ¹ H NMR (300MHz, CDCl ₃ ) δ: 0.87～0.91 (3H, t), 1.27 (18H, s), 1.56～1.60 (2H, t), 2.53～2.57 (1H, t), 3.43～3.47(2H, t), 3.56～3.76(36H, m).

The specific results of the chromatogram results of DD-PEG9-OH are shown in Table 3. The chromatogram is shown in Figure 10. Figure 11 is the chromatogram of the 1% reference substance of DD-PEG9-OH. The mass spectrum results are shown in Figure 12. MS (ESI+): m/z+H=582.8.

Table 3 Chromatogram results of DD-PEG9-OH

4. Chemical synthesis, separation, purification and characterization of DD-PEG10-OH

Add HO-PEG10-OH (18g, 40mmol, 2eq) to tetrahydrofuran (144mL, 8V), add sodium hydride (60%, 1.2g, 1.5eq) with stirring and react for 20min, 1-bromododecane (5g) , 20 mmol) was dissolved in tetrahydrofuran (10 mL) and added dropwise to the system. After the drop was completed, the system was heated to 80°C and reacted overnight. TLC monitors the reaction of the raw materials. Add saturated NH ₄ Cl solution to adjust the pH of the system <7. Filter the salt and spin dry the tetrahydrofuran. Add water (100ml) to dilute, extract with dichloromethane (3*100ml), separate the liquids, and combine the organic phases. The water was dried over Na ₂ SO ₄ , filtered, and concentrated to dryness. Column chromatography was carried out with MeOH:DCM=1:10, and 14 g of light yellow solid was eluted, with a yield of 55.8%.

The NMR results are shown in Figure 14. ¹ H NMR (300MHz, CDCl ₃ ) δ: 5.544 (1H, t), 3.518～3.366 (40H, m), 1.477～1.455 (2H, t), 1.249 (18H, s) ,0.863(3H,t).

The specific results of the chromatogram results of DD-PEG10-OH are shown in Table 4. The chromatogram is shown in Figure 15. Figure 16 is the chromatogram of the 1% reference substance of DD-PEG10-OH. The mass spectrum results are shown in Figure 17. MS (ESI+): m/z+H=628.1.

Table 4 Chromatogram results of DD-PEG10-OH

5.Chemical synthesis, isolation, purification and characterization of DD-PEG11-OH

Add HO-PEG11-OH (20.6g, 40mmol, 2eq) to tetrahydrofuran (164mL, 8V), add sodium hydride (60%, 1.2g, 1.5eq) with stirring and react for 20min, 1-bromododecane (5g) , 20 mmol) was dissolved in tetrahydrofuran (10 mL) and added dropwise to the system. After the drop was completed, the system was heated to 80°C and reacted overnight. TLC monitors the reaction of the raw materials. Add saturated NH ₄ Cl solution to adjust the pH of the system <7. Filter the salt and spin dry the tetrahydrofuran. Add water (100ml) to dilute, extract with dichloromethane (3*100ml), separate the liquids, and combine the organic phases. The water was dried over Na ₂ SO ₄ , filtered, concentrated to dryness, and subjected to column chromatography with methanol:dichloromethane = 1:10 to obtain 1.2g of pure pale yellow solid product and 14g of crude pale yellow solid product. The yield of pure product was 4.5%.

The NMR results are shown in Figure 18. ¹ H NMR (300MHz, CDCl ₃ ) δ: 3.894~3.569 (42H, m), 3.48~3.397 (2H, m), 2.689 (1H, s), 1.609 (2H, t) ,1.586(18H,s)1.563(3H,t).

The specific results of the chromatogram results of DD-PEG11-OH are shown in Table 5. The chromatogram is shown in Figure 19. Figure 20 is the chromatogram of the 1% reference substance of DD-PEG10-OH. The mass spectrum results are shown in Figure 21. MS (ESI+): m/z+H=672.3.

Table 5 Chromatogram results of DD-PEG11-OH

3. Synthesis, separation, purification and characterization of DD-PEG12-OH

Add sodium hydride (368 mg, 9.15 mmol) to a 500 ml single-neck bottle containing dimethyl sulfoxide (200 ml) at room temperature, stir for 30 minutes, then slowly add dodecaethylene glycol (20 g, 36.6 mmol) under nitrogen protection, and stir at room temperature. 2h. Under nitrogen protection, 1-bromododecane (2.28g, 9.15mmol) was slowly added into the reaction flask, and then the reaction was raised to 90°C and stirred for 18h. The reaction was brought to room temperature, water (400 ml) was added, and extracted with ethyl acetate (100 ml) for a total of 3 times. The organic phases were combined and washed with saturated sodium chloride solution (100 ml). The organic phase was dried with anhydrous Na ₂ SO ₄ , filter, concentrate to dryness, dissolve dichloromethane and load the sample, forward separation, the separation gradient is: dichloromethane: methanol (0% ~ 4%), TLC spot plate determines the product point, the product fractions are combined and concentrated to obtain 1.64g white Solid, yield 25.1%.

The NMR results are shown in Figure 10. ¹ HNMR (300MHz, CDCl ₃ ) δ: 0.87～0.92 (3H, t), 1.27 (18H, s), 1.56～1.61 (2H, t), 2.54～2.58 (1H, t) ), 3.43～3.48(2H, t), 3.57～3.76(48H, m).

The specific results of the chromatogram results of DD-PEG12-OH are shown in Table 6. The chromatogram is shown in Figure 23. Figure 24 is the chromatogram of the 1% reference substance of DD-PEG12-OH. The mass spectrum results are shown in Figure 26. MS (ESI+):m/z 715.0.

Table 6 Chromatogram results of DD-PEG12-OH

1.1.4 Synthesis, isolation, purification and characterization of DD-PEG24-OH

Add HO-PEG24-OH (21.4g, 20mmol, 2eq) to tetrahydrofuran (170mL, 8V), add sodium hydride (60%, 600mg, 1.5eq) with stirring and react for 10min, 1-bromododecane (2.5 g, 10 mmol) dissolved in tetrahydrofuran (10 mL) and added dropwise to the system. After the dripping was completed, the system was heated to 80°C and allowed to react overnight. TLC monitors the reaction of the raw materials. Add saturated NH ₄ Cl solution to adjust the pH of the system to <7. Filter the salt and spin dry the tetrahydrofuran. Add water (100ml) to dilute, extract 3 times with dichloromethane (100ml), separate the liquids, and combine the organic phases. Dry over anhydrous Na ₂ SO ₄ , filter, concentrate to dryness, and perform column chromatography with methanol: dichloromethane = 1:10. 2g of light yellow solid is eluted, with a yield of 8%.

The NMR results are shown in Figure 27. ¹ HNMR (300MHz, CDCl ₃ ) δ: 3.733~3.765 (1H, J=5.4Hz; t), 3.264~3.652 (96H, m), 1.454~1.497 (2H, J=6.6 Hz; t), 1.249 (18H, s), 0.839~0.884 (3H, J=6.6Hz; t).

Using the self-control method, the purity of DD-PEG24-OH is 98.06%. The specific results of the chromatogram of DD-PEG24-OH are shown in Table 7. The chromatogram is shown in Figure 28. Figure 29 is 1% of DD-PEG24-OH. The chromatogram and mass spectrum results of the reference substance are shown in Figure 30, MS (ESI+): 1/2m/z+H=622.6.

Table 7 Chromatogram results of DD-PEG24-OH

Example 2 In vitro experiments comparing the cytotoxicity of single molecular weight polidocanols with different chain lengths

Comparisons were made between polidocanol (commercially available as a mixture of different molecular weights) and single molecular weight polidocanols DD-PEG8-OH, DD-PEG9-OH, DD-PEG10-OH and DD. -PEG11-OH, diluted cytotoxicity.

2.1 Experimental steps:

HUVEC (Human Umbilical Vein Endothelial Cells) seed plates were used to dilute the test products in concentration gradients. After diluting to the same concentration, they were added to 96-well plates seeded with cells. After incubation for 24 to 72 hours, CCK8 was used for detection. Number of viable cells and calculate _IC50 value.

Specific cell culture steps:

(1) Passage HUVEC (human umbilical vein endothelial cells) continuously for 2 to 3 times to keep the cells in the logarithmic growth phase.

(2) Prepare cell suspension

Take HUVEC (human umbilical vein endothelial cells) from 10 to 30ml to a 50ml sterile centrifuge tube, and centrifuge at 1000r/min Collect the cells for 5 minutes, discard the supernatant, wash once with phosphate buffer saline (DPBS), resuspend in basic culture medium, count with a cell counting board, adjust the cell concentration to approximately 1.0×10 ⁴ cells/ml, and set aside.

(3) Cell seed plate

Use a multi-channel pipette to pipette the cell suspension until the cells are fully mixed, then add them to a 96-well plate.

(4) Test sample dilution: Take polidocanol (purchased from Siegfried Hameln GmbH, Germany, batch number OK13103) and prepared in Example 1: DD-PEG4-OH, DD-PEG8-OH, DD-PEG9-OH, DD-PEG10-OH, DD-PEG11-OH, DD-PEG24-OH, and PBS solution (containing ethanol with a mass concentration of 4%) were used to prepare test sample mother solutions of appropriate concentrations.

(5) Gradiently dilute the test sample mother solution to the set concentration (concentrations are: 0.001524, 0.004572, 0.01372, 0.04115, 0.1235, 0.3704, 1.111, 3.333, 10.00mM), and add them to the 96 in which the cells have been seeded. orifice plate.

(6) Culture: place at 37±1°C and 5±1% _CO2 for 18 to 24 hours.

(7) Read the plate: Vibrate and mix, then compare the color on a microplate reader. The experimental wavelength is 570nm and the reference wavelength is 630nm.

In this example, the diluted concentrations of the test samples in each group are equal.

Experimental results:

HUVEC (human umbilical vein endothelial cells) were used to test the IC ₅₀ values of different types of single molecular weight polidocanol samples. The results are shown in Table 8. The IC ₅₀ values of the DD-PEG8-OH, DD-PEG9-OH, DD-PEG10-OH, DD-PEG11-OH, and DD-PEG12-OH groups were all smaller than those of the polidocanol group, while the DD-PEG4-OH, DD -The IC ₅₀ value of PEG24-OH is greater than that of the polidocanol group, indicating that the PEG length in a single molecular weight polidocanol will affect its cytotoxicity if it is too long or too short, and the cytotoxicity is directly proportional to the efficacy of polidocanol. , indicating that when a single molecular weight polidocanol contains 8, 9, 10, 11 or 12 PEGs, its efficacy is better than that of the prior art.

Table 8 IC ₅₀ values of single molecular weight polidocanol with different chain lengths for HUVEC

Example 3 Comparison of Foaming Efficiency of Single Molecular Weight Polydocanol with Different Chain Lengths

3.1 Experimental methods:

3.1.1 Weigh appropriate amounts of single molecular weight polidocanol samples (DD-PEG4-OH, DD-PEG8-OH, DD-PEG9-OH, DD-PEG10-OH, DD-PEG11 prepared in Example 1). -OH, DD-PEG12-OH, DD-PEG24-OH single molecular weight polidocanol samples) and equal amounts of polidocanol samples (containing polidocanol in a mixture state of different molecular weights, purchased from Germany Siegfried Hameln GmbH, batch number OK13103), use PBS solution (which contains ethanol with a mass concentration of 4%) to prepare a 1% polidocanol solution, and let it stand until the foam disappears.

3.1.2 Use a 2.5ml syringe to absorb 0.5ml of polidocanol solution with a concentration of 1%, and another syringe to absorb 1.5ml of air. Connect the three-way tubes respectively, and then pump 20 times until a uniform solution is formed. Foam, collect all the foam into a syringe, adjust the volume to 2ml, and start observing.

3.1.3 Let it stand and observe the foam state, and record the following parameters:

(1) The time when liquid begins to separate:

Foam drainage time (FDT) is the length of time the hardener liquid is visible.

(2) The time it takes for half of the foam volume to disappear

Foam half-life (FHT) is the time it takes for the foam emptying rate to reach 50%;

(3) The time required for the bubble to completely disappear:

Foam coalescence time (FCT) is the length of time the hardener foam is visible.

P4 represents the single molecular weight polydocanol of DD-PEG4-OH, P8 represents the single molecular weight polydocanol of DD-PEG8-OH, and P9 represents the single molecular weight polydocanol of DD-PEG9-OH. In this way analogy.

The polidocanol in this embodiment is a mixture of polidocanols containing different molecular weights and not a single molecular weight.

3.2 Result analysis:

3.2.1 Time when liquid begins to separate:

Foam drainage time (FDT) is the length of time the hardener liquid is visible. The results are shown in Figure 31. The abscissa is the sample of each molecular weight, and the ordinate is the time when the liquid appears (unit s). The P4 sample (that is, DD-PEG4-OH) is difficult to form foam. After other samples form uniform foam, Liquid will precipitate within 3 to 8 seconds, and the precipitation time of P10 sample is relatively slow.

3.2.2 Time for half of the foam volume to disappear

Foam half-life time (FHT) is the time it takes for the foam emptying rate to be 50%. As shown in Figure 32, it is difficult for the P4 sample to form foam, and even if a small amount of foam is formed, it will disappear in a very short time. Other samples can form uniform foam after 20 pumps, among which the foam half-life of P8 to P24 is more than 120s.

3.2.3 The time required for the bubble to completely disappear:

Foam coalescence time (FCT) is the length of time the hardener foam is visible. As shown in Figure 33, the time required for the foam of the P24 sample to completely disappear is shorter, about 4.25 minutes. The time required for the foam of P8, P9, P10 and polidocanol to completely disappear ranged from 13.75 to 19.5 minutes.

Example 4 In vivo experiments to evaluate the venous sclerosis effect of single molecular weight polidocanol with different chain lengths

Compare the venous sclerosis effects of the mixtures of DD-PEG8-OH, DD-PEG9-OH, DD-PEG10-OH, DD-PEG11-OH and polidocanol prepared in Example 1.

Specifically, healthy New Zealand rabbits were averagely divided into the following groups: negative control, positive control (low), positive control (high), P8, P9 (low), P9 (high), P10, and P11. Prepare the samples shown in Table 9 into injectable dosage forms. See 4.1.1 for the preparation method. The effect of a single administration of the above preparation on venous sclerosis of rabbit ears was observed, and the results are shown in Figures 34 to 37.

Table 9 Samples of venous sclerosis experiments of single molecular weight polidocanol with different chain lengths

4.1.1 Preparation of foam preparation

Foamed formulations of the samples shown in Table 9 were prepared.

Preparation:

(1) Connect two syringes (the minimum scale is 0.25ml, easy for injection) to the tee tube. One of them sucks 1 volume of sample, and the other sucks 3 volumes of air. Then pump back and forth 40 times until a uniform foam forms.

(2) Suck all the foam into one syringe, remove it from the tee tube and prepare for injection; empty the other syringe and keep it on the tee tube.

(3) Foam injection must be completed within 1 minute. After injecting an animal, the syringe needs to be connected to the tee tube again and aspirated repeatedly until a uniform foam is formed. Then repeat step (2).

4.1.2 Animal drug administration process

After adaptive feeding for 7 days, healthy New Zealand rabbits were divided into negative control, positive control (low), positive control (high), P8, P9 (low), P9 (high), P10, and P11 groups on average. Insert the needle into the distal end of the marginal ear vein and slowly inject the samples shown in Table 9. The injection process must be completed within 1 minute.

4.1.3 Detection indicators

Photographs of rabbit ear blood vessels were collected before administration, 7 days, 14 days and 21 days after administration to observe phenomena such as vascular hardening and blockage.

4.2 Experimental results:

4.2.1. Results after 14 days of administration through the marginal ear vein of rabbits

Figure 34 shows the appearance of the veins of some rabbits 14 days after intravenous injection of different types of polidocanol into the ear margins of some rabbits in this example (p8 group, P9 low-dose group, positive control low-dose group). After 14 days of administration, some Blood vessels are blocked. From the appearance, the color is darker and lighter, and the blood flow of some uninjected blood vessels becomes larger. All rabbit ear blood vessels were scored, valid as 1 and invalid as 0.

The calculation rule for effective efficiency is: effective rate (%) = rabbit ear score / total number of rabbit ears * 100% (calculated based on the number of rabbit ears)

The effective rates of venous sclerosis 14 days after intravenous injection of different types of polidocanol into the rabbit ear margin are shown in Table 10 and Figure 35. From Table 10 and Figure 35, it can be seen that the P8 group, P10 group, P1 group, P9 high-dose group and P9 The effective rates in the low-dose group were all above 83.33%, not lower than the positive control low-dose group.

Table 10 Effective rates of venous sclerosis 14 days after intravenous injection of different types of polidocanol into rabbit ear margins

4.2.2. Results after 21 days of administration through the marginal ear vein of rabbits

Figure 36 shows the appearance of veins 21 days after intravenous injection of different types of polidocanol into some rabbit ear margins in this example (p8 group, p9 low group, positive control low group). After 21 days of administration, some blood vessels in the rabbit ear Completely blocked. From the appearance, the color of some blood vessels becomes lighter or gradually disappears, and the blood flow of uninjected blood vessels increases.

All rabbit ear blood vessels were scored, valid as 1 and invalid as 0. See 4.2.1 for calculation rules.

The results in Table 11 and Figure 37 show that the effective rates of P8 group, P10 group, P11 group, P9 high-dose group and P9 low-dose group are all higher than the positive control low-dose group, but the overall venous sclerosis effective rate in Table 11 is lower than Table 10 and Figure 35, this is because after polidocanol destroys the vascular endothelium, fibrosis will gradually occur in the blood vessels. On the 21st day, some blood vessels will gradually become blocked until they are blocked, and some blood vessels will open again.

Table 11 Effective rates of venous sclerosis after 21 days of intravenous injection of different types of polidocanol into rabbit ear margins

4.3. Venous pathological section results 14 days after intravenous injection of different types of polidocanol into rabbit ear margins

4.3.1 Pathological changes in blood vessels of animals in each group

Observe the blood vessels of the negative control group, positive high-dose group, positive low-dose group, P8 group, P10 group, P11 group, P9 high-dose group and P9 low-dose group after 14 days of administration to the marginal ear vein of rabbits in 4.2.1. Structural changes in each layer of the wall and inflammatory response, thrombosis, vascular fibrosis, etc.

4.3.2 Assessment of pathological damage degree:

According to the scoring system proposed by AlGhamdi K M ^[1] and others, the pathological changes under venoscopy in each group were scored from aspects such as inflammatory response, vascular proliferation, vascular lumen stenosis, and fibrosis to evaluate the degree of pathological damage to the blood vessels. Score is 10 points.

The results are shown in Table 12. The pathological scores of rabbit ear blood vessels in the positive control group were significantly higher than those in the negative control group. Groups P8, P10, The pathological scores of the P11 group, P9 high-dose group and P9 low-dose group were higher than those of the positive low-dose control group.

Table 12 Mean table of rabbit ear vascular pathological scores in each group

The specific results of the pathological changes in the marginal ear veins of rabbits after 14 days of administration are shown in Figure 38. Under a low-power microscope (magnification 20×) in the negative control group, the lumen of the blood vessels was unobstructed, with no obvious stenosis, and the fibrous connective tissue around the blood vessels was significantly loose and edematous. , a small amount of inflammatory cell infiltration can be seen in the connective tissue. In the negative control group, under a high-power microscope (magnification 200×), vascular endothelial cells and adventitial fibrous connective tissue were visible. The vascular endothelial cells were arranged neatly and tightly; a small amount of neutrophil infiltration was seen in the fibrous connective tissue around the blood vessels.

P8, P9 low-dose groups, P9 high-dose groups, P10, P11 and the positive control group (polidocanol group) high- and low-dose groups all have pathological changes in blood vessels. The specific description is as follows: Under low magnification, the blood vessels The tissue structure of each layer of the wall is unclear and the blood vessel lumen is occluded. Under high magnification, a large number of fibroblast proliferations can be seen in the lumen, and red blood cells are scattered among local fibroblasts; new capillaries and macrophages that phagocytose hemosiderin can be seen locally. The above results show that the single molecular weight polidocanol of the present invention can effectively fibrosis the vascular endothelium and further seal the vascular lumen.

Reference non-patent literature: [1] Alghamdi K.M., Ashour A.E., RikabiA.C., et al.Phenol as a novel sclerosing agent: A safety and efficacy study on experimental animals[J].Saudi PharmJ.2014; 22(1) :71-8

Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention, but not to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: It is still possible to modify the technical solutions recorded in the foregoing embodiments, or to equivalently replace some or all of the technical features; and these modifications or substitutions do not deviate from the essence of the corresponding technical solutions from the technical solutions of the embodiments of the present invention. range.

Claims (12)

1. The application of polydocanol with a single molecular weight in the preparation of disease treatment drugs is characterized in that the polydocanol is used as a sclerosing agent and/or hemostatic agent for treatment.
2. The application according to claim 1, wherein the diseases include cystic diseases, vascular related diseases, tumors, obstetric and gynecological diseases, inflammation and/or pain;

   Preferably, the vascular related diseases include varicose veins, vascular malformations and/or hemorrhage;

   Preferably, the obstetric and gynecological diseases include uterine submucosal fibroids and cesarean scar pregnancy;

   Preferably, the cystic disease includes abdominal organ cysts, body surface cysts, lymphoceles, thyroid cysts, bone cysts, digital mucocele and/or pelvic encapsulated effusion;

   Preferably, the tumors include benign or malignant solid tumors;

   Preferably, the inflammation includes infectious inflammation and/or non-infectious inflammation;

   Preferably, the pain includes chronic pain, acute pain and/or cancer pain.
3. The application according to claim 2, wherein the tumors include: infantile hemangioma, congenital hemangioma, plexiform hemangioma, spindle cell hemangioma, epithelioid hemangioma, pyogenic granuloma, cardinal hemangioma, Posiform hemangioendothelioma, reticular hemangioendothelioma, papillary intralymphatic hemangioendothelioma, composite hemangioendothelioma, Kaposi's sarcoma, angiosarcoma, epithelioid hemangioendothelioma, uterine fibroids and/or abdominal viscera malignant tumors;

   The pain includes: muscle and soft tissue pain, bone and joint pain, and/or visceral pain;

   The inflammation includes: tendonitis, tendinitis, tendinosis, peritendinitis, spondylitis, sacroiliitis, gouty arthritis, myofasciitis, tenosynovitis and/or frozen shoulder;

   The varicose veins include: varicose veins of the lower limbs, esophageal and gastric varices, varicocele, abdominal wall varicose veins and/or hemorrhoids;

   The bleeding includes: gastrointestinal bleeding, bladder bleeding, bleeding caused by early, middle or late cancer;

   The abdominal organ cysts include: liver cysts, renal cysts, pararenal pelvic cysts, pancreatic cysts, endometriosis cysts and/or ovarian chocolate cysts;

   The body surface cysts include: skin cysts and/or finger mucous cysts;

   The abdominal organ malignant tumors include liver cancer.
4. The application according to any one of claims 1 to 3, characterized in that the molecular structure of the single molecular weight polydocanol monomer is:
   

   Where n is an integer, 4≤n≤24;

   Preferably, n is an integer of 8≤n≤15;

   More preferably, n is 8, 9, 10 or 11.
5. A polydocanol with a single molecular weight, characterized in that the polydocanol with a single molecular weight is prepared by the following preparation method: react polyethylene glycol with a single molecular weight and 1-halododecane, separate, The product was purified to obtain polidocanol of a single molecular weight.
6. Polydocanol as claimed in claim 5, characterized in that the preparation method includes the following steps:

   S1: Mix single molecular weight polyethylene glycol with catalyst and organic solvent, add 1-halododecane dropwise, raise the temperature to the preset temperature, and keep the reaction warm;

   S2: After the reaction is completed, cool down the reactant. After it reaches room temperature, add water to dilute, extract, and separate the liquids;

   S3: Filter, concentrate, dry, separate and purify the organic phase to obtain the single molecular weight polydocanol.
7. Polydocanol according to claim 6, characterized in that the catalyst is one or more of sodium hydroxide, sodium hydride, potassium hydroxide, potassium hydride, potassium tert-butoxide or sodium tert-butoxide, Preference is given to sodium hydroxide or sodium hydride.
8. Polydocanol according to claim 6, characterized in that the organic solvent is selected from: ethanol, methanol, isopropyl alcohol, tetrahydrofuran, acetone, acetonitrile, toluene, xylene, n-hexane, cyclohexane, isopropanol, At least one of octane, n-pentane, petroleum ether, dimethyl sulfoxide, N,N-dimethylformamide dichloromethane, and diethoxymethane; preferably tetrahydrofuran, dimethyl sulfoxide, At least one of acetonitrile, toluene, and N,N-dimethylformamide.
9. Polydocanol according to claim 6, characterized in that the 1-halododecane is selected from 1-chlorododecane, 1-bromododecane or 1-iodododecane, preferably 1-bromododecane.
10. The polidocanol according to claim 6, characterized in that: the preset temperature is 80-95°C, preferably 80-90°C;

    The time of the insulation reaction is 12 to 24 hours. Preferably, the time of the insulation reaction is 16 to 24 hours. More preferably, the time of the insulation reaction is 16 to 20 hours;

    The extraction solvent is one or a combination of more of diethyl ether, ethyl acetate, chloroform, methylene chloride, toluene, and methyl tert-butyl ether. Preferably, the extraction solvent is ethyl acetate.
11. Polydocanol according to claim 6, characterized in that the separation and purification is carried out by chromatography; preferably column chromatography, and the eluent used in the column chromatography is selected from: ethyl acetate, One or more of methanol, ethanol, n-hexane, petroleum ether or methylene chloride.
12. A polidocanol preparation, characterized in that it includes the single molecular weight polidocanol or a pharmaceutically acceptable salt thereof according to any one of claims 5 to 11 and pharmaceutically acceptable auxiliary materials; preferably, The preparation is an injection. More preferably, the injection is a stock solution or a foaming preparation. More preferably, the preparation is a foaming preparation.