WO2023280239A1 - 绿原酸在制备治疗中枢神经系统肿瘤的药物中的用途 - Google Patents

绿原酸在制备治疗中枢神经系统肿瘤的药物中的用途 Download PDF

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WO2023280239A1
WO2023280239A1 PCT/CN2022/104224 CN2022104224W WO2023280239A1 WO 2023280239 A1 WO2023280239 A1 WO 2023280239A1 CN 2022104224 W CN2022104224 W CN 2022104224W WO 2023280239 A1 WO2023280239 A1 WO 2023280239A1
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chlorogenic acid
nervous system
central nervous
lymphoma
preparation
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French (fr)
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张洁
李文斌
黄望
张雅
张飞
徐敏
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四川九章生物科技有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
    • A61K31/216—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acids having aromatic rings, e.g. benactizyne, clofibrate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

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  • the invention belongs to the field of medicine, and in particular relates to a new application of chlorogenic acid in the preparation of medicines for treating central nervous system tumors.
  • Central nervous system lymphoma is a central nervous system tumor with a low incidence, accounting for 1% to 3% of central nervous system tumors. With the application of immunosuppressants, the incidence of the disease has been on the rise in recent years. Central nervous system lymphoma includes primary central nervous system lymphoma and secondary lymphoma in which systemic lymphoma invades the central nervous system. Primary central nervous system lymphoma, also known as primary central nervous system lymphoma (PCNSL), accounts for about 8% of central nervous system lymphoma.
  • PCNSL primary central nervous system lymphoma
  • Non-Hodgkin's lymphoma is a relatively rare type of non-Hodgkin's lymphoma (non-Hodgkin's lymphoma) that originates in the central nervous system (including brain, eyes, spinal cord, meninges, etc.)
  • Hodgkin lymphoma (NHL) accounts for about 2% to 4% of all intracranial tumors and 4% to 6% of all non-Hodgkin lymphoma (NHL). In recent years, its incidence has gradually increased.
  • B-NHL B-cell non-Hodgkin's lymphoma
  • Diffuse large B-cell lymphoma is the most common aggressive B-cell non-Hodgkin's lymphoma Lymphoma (NHL) is the most common type, accounting for about 30% to 40% of adult NHL.
  • DLBCL the current comprehensive treatment scheme based on high-dose methotrexate (HD-MTX) is the main one.
  • HD-MTX high-dose methotrexate
  • Chlorogenic acid also known as coffee tannin (Chlorogenic acid, CHA for short), is an organic acid widely present in honeysuckle, leaves of Eucommia ulmoides, sunflower seeds, capillary and blue potted flowers, etc. It has antioxidant, Antibacterial, antiviral, hypoglycemic, lipid-lowering, blood pressure-lowering and immune regulation and other pharmacological effects.
  • the Chinese patent application with the application number CN201510079639.8 discloses the use of chlorogenic acid in the preparation of drugs for the prevention and treatment of primary cutaneous T-cell lymphoma. The application proves through experiments that chlorogenic acid can activate CD4 T lymphocytes and CD8 T lymphocytes.
  • Lymphocytes and target CD4T lymphocytes and CD8T lymphocytes to prevent and treat primary cutaneous T-cell lymphoma.
  • primary cutaneous T-cell lymphoma is not a tumor of the central nervous system, and its etiology, location and treatment mechanism are quite different from those of central nervous system tumors. There is no report on the use of chlorogenic acid to treat central nervous system tumors.
  • the purpose of the present invention is to provide a new application of chlorogenic acid in the preparation of drugs for preventing and/or treating central nervous system tumors.
  • the invention provides the use of chlorogenic acid in the preparation of medicines for preventing and/or treating central nervous system tumors.
  • the central nervous system tumor is central nervous system lymphoma.
  • central nervous system lymphoma is primary central nervous system lymphoma.
  • the primary central nervous system lymphoma is B-cell non-Hodgkin's lymphoma.
  • B-cell non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.
  • the diffuse large B-cell lymphoma is diffuse large B-cell lymphoma in the brain, eye or spinal cord.
  • the medicine is a pharmaceutical preparation prepared by taking chlorogenic acid as an active ingredient and adding pharmaceutically acceptable auxiliary materials.
  • the pharmaceutical preparation is an oral preparation.
  • the pharmaceutical preparation is an injection preparation.
  • the unit preparation of the pharmaceutical preparation contains 0.5-5.5 mg of chlorogenic acid.
  • the unit preparation of the pharmaceutical preparation contains 1.1-3.3 mg of chlorogenic acid, preferably 3.3 mg.
  • the present invention also provides a medicine for treating primary central nervous system lymphoma, which is a medicine prepared by taking chlorogenic acid as an active ingredient and adding pharmaceutically acceptable auxiliary materials; the unit preparation of the medicine is Contains 0.5-5.5 mg of chlorogenic acid, preferably 1.1-3.3 mg, more preferably 3.3 mg.
  • the present invention finds for the first time that chlorogenic acid can effectively treat diffuse large B-cell lymphoma, and provides a new and safe option for the clinical treatment of central nervous system tumors, especially diffuse large B-cell lymphoma. It has a good application prospect.
  • Figure 1 shows the growth curves of mice with brain diffuse large B-cell lymphoma in each administration group.
  • Fig. 2 is the tumor inhibition rate of each administration group to mice.
  • Figure 3 shows the growth curves of orbital diffuse large B-cell lymphoma mice in each administration group.
  • Fig. 4 is the tumor inhibition rate of each administration group to mice.
  • the raw materials and equipment used in the present invention are known products obtained by purchasing commercially available products.
  • Embodiment 1 Chlorogenic acid oral preparation prescription of the present invention
  • Preparation method Weigh chlorogenic acid according to the prescription, and aseptically pack it into powder.
  • Preparation method Weigh chlorogenic acid, fillers, and binders according to the prescription, granulate, granulate, and pack into granules.
  • Preparation method Weigh chlorogenic acid, filler and binder according to the prescription, granulate, granulate, add lubricant, compress into tablets to obtain tablets.
  • the filler is one or more of mannitol, lactose, starch, microcrystalline cellulose, and dextrin;
  • the binder is one of sodium carboxymethyl cellulose and PVP (polyvinylpyrrolidone) or two;
  • the lubricant is one or more of magnesium stearate, talcum powder, and micronized silica gel.
  • Embodiment 2 Prescription of chlorogenic acid injection preparation of the present invention
  • Preparation method (1) Aseptically weigh chlorogenic acid according to the prescription, mix evenly, and aseptically pack into powder injection.
  • Preparation method (2) Weigh chlorogenic acid according to the prescription, dissolve it in water for injection, filter and sterilize, and freeze-dry to obtain a freeze-dried powder injection.
  • Preparation method Weigh chlorogenic acid, scaffolding agent, and antioxidant according to the prescription, dissolve them in water for injection, filter and sterilize, and freeze-dry (freeze-drying conditions: pre-freezing: temperature ⁇ -40°C, normal pressure, drying time 2-4h , freeze; primary drying: temperature ⁇ -13°C, negative pressure, drying time ⁇ 12h, fully dry; secondary drying, temperature 20-30°C, negative pressure, drying time ⁇ 2h), to obtain lyophilized powder injection.
  • freeze-drying conditions pre-freezing: temperature ⁇ -40°C, normal pressure, drying time 2-4h , freeze; primary drying: temperature ⁇ -13°C, negative pressure, drying time ⁇ 12h, fully dry; secondary drying, temperature 20-30°C, negative pressure, drying time ⁇ 2h
  • the scaffolding agent is one or more of mannitol, lactose, and glucose; the antioxidant is one or more of sodium bisulfite, vitamins, glutathione, and folic acid.
  • Test Example 1 In vitro test of chlorogenic acid in treating central nervous system lymphoma
  • Diffuse large B-cell lymphoma cell line pfeiffer was purchased from ATCC in the United States and subcultured in RPMI 1640 medium.
  • methotrexate for injection provided by Jiangsu Hengrui Medicine Co., Ltd.
  • Test drug chlorogenic acid raw material drug.
  • the above two drugs were prepared to the required concentration with serum-free RPMI 1640 medium, then sterilized by filtration through a 0.22 ⁇ m microporous membrane, and stored in a 4°C refrigerator for later use.
  • cryopreservation tube containing the cells into a 37°C water bath to heat, shake quickly to melt it as soon as possible, put the thawed cell suspension into a 10ml centrifuge tube, centrifuge at a low speed of 1000r/min for 5-10min, discard the supernatant, Add an appropriate amount of fresh medium, blow and beat the cells gently, mix well and then centrifuge again (1000r/min) for 5min to minimize the DMSO component in the medium. After centrifuging the cells again, discard the supernatant, add fresh medium and 10% FBS, pipette the cells and medium to mix evenly, put them into a culture bottle, and culture them in an incubator with a constant temperature of 37°C and 5% CO 2 .
  • the experiment was divided into 8 groups: blank group, negative control group, positive control group and 5 test drug groups.
  • Blank group only adding medium, no need to inoculate pfeiffer cells
  • Negative control group add medium and inoculate pfeiffer cells
  • Positive control group 10 ⁇ g/ml methotrexate was cultured with pfeiffer cells;
  • Test drug group the test drug is chlorogenic acid, the experimental concentration is 5 ⁇ g/ml, 10 ⁇ g/ml, 30 ⁇ g/ml, 50 ⁇ g/ml, 100 ⁇ g/ml, and it is cultured with pfeiffer cells.
  • drugs were added in groups, and the cells were incubated at a constant temperature of 37°C and 5% CO 2 for 48 hours.
  • the dosage of each well was 100 ⁇ l, the blank group was not inoculated with cells, and 100 ⁇ l of the same amount of medium was added, and the negative control group was added with 100 ⁇ l of the same amount of medium on the basis of inoculation of cells, and each group had 3 replicate wells.
  • the OD value was measured using an enzyme-linked immunosorbent assay instrument, zeroed with a blank control, and the wavelength used was 490 nm.
  • the growth inhibition rate of tumor cells was calculated according to the following formula:
  • GI (growth inhibition rate) (1-OD drug group/OD negative control group) ⁇ 100%.
  • the cell morphology was observed under an inverted microscope.
  • the positive control group and the chlorogenic acid test drug group had certain effects on cell growth, and some cells became round, shed, and suspended.
  • the chlorogenic acid test drug group had a significant inhibitory effect on the growth of pfeiffer cancer cell line at a concentration of 50-100 ⁇ g/ml, and a large number of cells became round, shed, and suspended.
  • Table 1 shows the inhibition rate of each administration group on pfeiffer cancer cells.
  • the chlorogenic acid test drug group has obvious tumor inhibitory effect on pfeiffer cancer cells, and when the concentration of chlorogenic acid is 50-100 ⁇ g/ml, the tumor inhibition rate is significant, as high as 71.68-72.97%; , the same dose (10 ⁇ g/ml) of chlorogenic acid has a higher inhibitory effect on pfeiffer cancer cells than the positive control drug methotrexate.
  • chlorogenic acid has obvious inhibitory effect on diffuse large B-cell lymphoma cell pfeiffer in vitro, and in this system, the inhibitory effect of chlorogenic acid concentration of 50-100 ⁇ g/ml is more significant.
  • Test drug 1 chlorogenic acid for injection, prescription ratio: chlorogenic acid, mannitol, sodium bisulfite (30:80:2);
  • Test drug 2 oral preparation of chlorogenic acid, prescription ratio: chlorogenic acid, filler (lactose), binder (sodium carboxymethyl cellulose) (1000:500:5);
  • chlorogenic acid preparations for injection weighed chlorogenic acid, mannitol, and sodium bisulfite according to the prescription ratio, dissolved in water for injection, sterilized by filtration, and freeze-dried (freeze-drying conditions: pre-freezing: temperature ⁇ -40°C, Atmospheric pressure, drying time 2-4h, freezing; primary drying: temperature ⁇ -13°C, negative pressure, drying time ⁇ 12h, fully dry; secondary drying, temperature 20-30°C, negative pressure, drying time ⁇ 2h), The labeled amount of chlorogenic acid was obtained as a freeze-dried powder injection of 30 mg/support.
  • the oral preparation of chlorogenic acid is weighed and mixed with chlorogenic acid, lactose and sodium carboxymethyl cellulose according to the ratio of the prescription, granulated, granulated, and subpackaged into granules.
  • Diffuse large B-cell lymphoma cell Ly8 was purchased from the Cancer Hospital affiliated to Fudan University. It was cultured in suspension in 10% calf serum DMEM complete culture medium, passaged, and grew stably. The cells were collected during the logarithmic growth phase of the cells, centrifuged and counted to dilute the cells to a concentration of 10 4 / ⁇ l for use. The amount of inoculated cells was 2 ⁇ 10 4 cells/only.
  • BALB/c mice 60 mice, half male and half female, aged 3 to 4 weeks, weighing 15 to 22 g, purchased from the Experimental Animal Management Center of West China Medical Center. Drinking water was carried out under sterile conditions.
  • mice were anesthetized by intraperitoneal injection of 0.2 ⁇ l of 1% pentobarbital sodium, fixed in a stereotaxic apparatus, and 2% iodine chlorhexidine solution was used to disinfect the skin of the skull.
  • the injection site of the syringe is 0.1 cm from the forehead of the mouse to the left or right side of the cranial midline, and 0.3 cm in front of the coronal suture.
  • the micro-injection syringe penetrates the mouse cranial skin and skull in turn, and enters the intracranial about 1 to 2 mm.
  • mice were kept at 30 °C for 2 h. After the end of anesthesia, place it at room temperature of 25°C, and observe the surface changes 24 hours after inoculation. Symptom observation: The top of the head on the injection side is raised, the body is thin, listless, and the activity is reduced.
  • mice after successful inoculation were randomly divided into 7 groups, 6 in each group, including:
  • Negative control group intraperitoneal injection of normal saline, once a day;
  • Injection administration group 1 intraperitoneal injection of test drug 1, once a day, 5 mg/kg/time;
  • Injection group 2 intraperitoneal injection of test drug 1, once a day, 10 mg/kg/time;
  • Injection administration group 3 intraperitoneal injection of test drug 1, once a day, 30 mg/kg/time;
  • Injection administration group 4 intraperitoneal injection of test drug 1, once a day, 50 mg/kg/time.
  • test drug groups were administered by intraperitoneal injection of 0.2 mL per 10 g of mouse body weight. It is used and formulated before daily administration, and administered for 14 days.
  • mice During the administration period, other adverse reactions such as tumor growth, food intake, and activity status of the mice were observed and recorded every other day. After 14 days of administration, the mice were killed by cervical dislocation, and the body weight and tumor size were measured and recorded according to the number before administration. Tumor inhibition rate (%) was calculated based on tumor weight. Body weight and tumor weight mean ⁇ standard deviation Express, and carry out t test between each administration group and negative control group, each administration group and positive control methotrexate group.
  • mice 60 BALB/c mice were inoculated with Ly8 cells to establish a xenograft tumor model.
  • a localized bulge was seen at the initial inoculation site, which disappeared in 2-3 days, and a small bulge began to grow in some BALB/c mice at the inoculation site after about 5 days; Afterwards, the bulge of the site gradually grew, and in about 14 days, the maximum diameter of the transplanted tumor grew to about 5mm.
  • a total of 45 BALB/c mice successfully established xenograft tumor models and continued to grow. The inoculation success rate was 75%.
  • mice after successful inoculation were selected, randomly divided into 7 groups, administered according to the test plan, and the body weight and tumor weight of the mice were recorded. The results are shown in Table 2 and Figure 1 and Figure 2.
  • the positive drug group, the 30mg/kg oral chlorogenic acid granule administration group, and the 10-30mg/kg chlorogenic acid injection dosage group had good therapeutic effects on diffuse large B-cell lymphoma in the brain. There was a significant difference between the negative control group (P ⁇ 0.05).
  • mice died in the negative control group 1 mouse died in the positive control group, oral granule group, 5mg/kg and 50mg/kg injection groups, and no mice in the 10-30mg/kg injection group death, indicating that the therapeutic effect of the effective dose of chlorogenic acid for injection group was good; in addition, after close observation, the mice in the negative control group showed lack of gloss in the fur, loss of appetite, and decreased activity.
  • the above symptoms became more and more obvious , the symptoms of mice in the effective dose treatment group were significantly improved, and there were no adverse reactions related to treatment.
  • chlorogenic acid has a good in vivo therapeutic effect on brain diffuse large B-cell lymphoma, and the injection dose of 10mg/kg ⁇ 30mg/kg has a higher tumor inhibition rate, and the injection dose of 30mg/kg Dosage works best.
  • mice According to "Pharmacological Experimental Methodology" edited by Xu Shuyun et al., calculated according to the dose per unit weight, the dose of mice is equivalent to 9.1 times that of humans. Therefore, it can be deduced that when chlorogenic acid is injected into the human body at a dosage of 3.3 mg/kg, it has the best therapeutic effect on patients with brain diffuse large B-cell lymphoma.
  • Test drug 1 chlorogenic acid for injection, prescription ratio: chlorogenic acid, mannitol, sodium bisulfite (30:80:2);
  • Test drug 2 oral preparation of chlorogenic acid, prescription ratio: chlorogenic acid, filler (lactose), binder (sodium carboxymethyl cellulose) (1000:500:5);
  • chlorogenic acid preparations for injection weighed chlorogenic acid, mannitol, and sodium bisulfite according to the prescription ratio, dissolved in water for injection, sterilized by filtration, and freeze-dried (freeze-drying conditions: pre-freezing: temperature ⁇ -40°C, Atmospheric pressure, drying time 2-4h, freezing; primary drying: temperature ⁇ -13°C, negative pressure, drying time ⁇ 12h, fully dry; secondary drying, temperature 20-30°C, negative pressure, drying time ⁇ 2h), The labeled amount of chlorogenic acid was obtained as a freeze-dried powder injection of 30 mg/support.
  • the oral preparation of chlorogenic acid is weighed and mixed with chlorogenic acid, lactose and sodium carboxymethyl cellulose according to the ratio of the prescription, granulated, granulated, and subpackaged into granules.
  • the diffuse large B-cell lymphoma cell line pfeiffer was purchased from ATCC, USA.
  • BALB/c mice 60 mice, half male and half female, aged 3 to 4 weeks, weighing 15 to 22 g, purchased from the Experimental Animal Management Center of West China Medical Center. Drinking water was carried out under sterile conditions.
  • Tumor cell suspension inoculation method was used. Before cell inoculation, all mice were irradiated with cesium 137 to further immunosuppress the mice to improve the success rate of inoculation, and tumor cells were inoculated within 6 hours after irradiation. After disinfection with 75% alcohol around the eyes, draw 0.1ml of tumor cell suspension with a 1ml empty needle, inject it into the orbit through the eyelid skin above the periorbital temporo, pull out the needle and press for 5min, observe no obvious abnormality after injection in nude mice, After inoculation, they were given autoclaved water containing gentamicin 100,000 U/L to prevent infection for one week, and 5% glucose was given orally and egg yolk feeding every day to enhance nutrition. The growth status of mice and local tumor growth were observed every day.
  • mice After successful inoculation, and randomly divide them into 7 groups, 6 in each group, including:
  • Negative control group intraperitoneal injection of normal saline, once a day;
  • Injection administration group 1 intraperitoneal injection of test drug 1, once a day, 5 mg/kg/time;
  • Injection group 2 intraperitoneal injection of test drug 1, once a day, 10 mg/kg/time;
  • Injection administration group 3 intraperitoneal injection of test drug 1, once a day, 30 mg/kg/time;
  • Injection administration group 4 intraperitoneal injection of test drug 1, once a day, 50 mg/kg/time.
  • test drug groups were administered by intraperitoneal injection of 0.2 mL per 10 g of mouse body weight. It is used and formulated before daily administration, and administered for 14 days.
  • mice During the administration period, other adverse reactions such as tumor growth, food intake, and activity status of the mice were observed and recorded every other day. After 14 days of administration, the mice were killed by cervical dislocation, and the body weight and tumor size were measured and recorded according to the number before administration. Tumor inhibition rate (%) was calculated based on tumor weight. Body weight and tumor weight mean ⁇ standard deviation Express, and carry out between each administration group and negative control group, the t test between each administration group and positive control methotrexate group and negative control group.
  • mice After inoculation in the right orbit of the mice, the eyeballs were slightly convex, and returned to normal on the second day.
  • 40 BALB/C mice had ocular symptoms, such as eyelids could not be opened, tears, and mild edema of the bulbar conjunctiva around the eyeballs.
  • 55 BALB/C nude mice had eye symptoms, tearing, obvious bulbar conjunctival edema around the eyeballs, no obvious exophthalmos, and the inoculation tumor formation rate was 91.67%.
  • mice after successful inoculation were selected, randomly divided into 7 groups, administered according to the test plan, and the body weight and tumor weight changes of the mice were recorded. The results are shown in Table 3 and Figure 3 and Figure 4.
  • chlorogenic acid has a good in vivo therapeutic effect on orbital diffuse large B-cell lymphoma, and the injection dosage of 10mg/kg ⁇ 30mg/kg has a higher tumor inhibition rate, and the dosage of 30mg/kg Works best. It can be deduced that when chlorogenic acid is injected into the human body at a dosage of 3.3mg/kg, the therapeutic effect on patients with orbital diffuse large B-cell lymphoma is the best.
  • the present invention provides the use of chlorogenic acid in the preparation of drugs for preventing and/or treating central nervous system tumors.
  • the present invention finds for the first time that chlorogenic acid can effectively treat diffuse large B-cell lymphoma, and provides a new and safe option for the clinical treatment of central nervous system tumors, especially diffuse large B-cell lymphoma. It has a good application prospect.

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Abstract

本发明提供了绿原酸在制备治疗中枢神经系统肿瘤的药物中的用途，属于医药领域。本发明首次发现绿原酸能够有效治疗弥漫性大B细胞淋巴瘤，为临床上中枢神经系统肿瘤，特别是弥漫性大B细胞淋巴瘤的治疗提供了一种新的、安全性高的选择，具有良好的应用前景。

Description

绿原酸在制备治疗中枢神经系统肿瘤的药物中的用途 技术领域
本发明属于医药领域，具体涉及绿原酸在制备治疗中枢神经系统肿瘤的药物中的新用途。
背景技术
中枢神经系统淋巴瘤是一种中枢神经系统肿瘤，该病发病率低，占中枢神经系统肿瘤的1％～3％。随着免疫抑制剂的应用，近年来该病发病率有上升趋势。中枢神经系统淋巴瘤包括原发中枢神经系统的淋巴瘤和全身淋巴瘤侵入中枢神经系统的继发性淋巴瘤。原发于中枢神经系统的淋巴瘤又称原发性中枢神经系统淋巴瘤(Primary central nervous system lymphoma，PCNSL)，约占中枢神经系统淋巴瘤的8％。
原发性中枢神经系统淋巴瘤是一种原发于中枢神经系统(包括脑、眼、脊髓、脑脊膜等)而无其他部位受累的一类较罕见的非霍奇金淋巴瘤(non-Hodgkin lymphoma，NHL)，约占所有颅内肿瘤的2％～4％，占所有非霍奇金淋巴瘤(NHL)的4％～6％，近年来其发病率呈逐渐增高趋势。多数原发于中枢神经系统的恶性淋巴瘤为B细胞非霍奇金淋巴瘤(B-NHL)，它可表现为弥漫性或多病灶性，临床上大多表现为偏瘫、行走不稳、视物模糊等神经症状，此外可表现为颅内压增高、癫痫发作、人格改变等。
弥漫性大B细胞淋巴瘤(diffuse large B celllymphoma，DLBCL)是一种最常见的侵袭性B细胞非霍奇金淋巴瘤，它是一种B细胞源性中、高度恶性肿瘤，是非霍奇金淋巴瘤(NHL)最常见的类型，约占成人NHL的30％～40％。对于DLBCL，目前以高剂量的甲氨蝶呤(high-dose methotrexate，HD-MTX)为基础的综合治疗方案为主。研究发现，化疗及放疗联合可显著提高对DLBCL的疗效，但HD-MTX联合全脑放疗(whole-brain radiotherapy，WBRT)会明显增加脑白质病的发病率，引起极大的长期神经毒性。此外，临床结果显示目前不到半数的患者通过标准化疗方案能够获得治愈，其余患者即使获得缓解也难免复发死亡。DLBCL难治的主要原因是细胞多药耐药基因过度表达，对化疗药物产生多药耐药。因此，亟需开发出一种毒副作用低、同时能够提高疗效的新药物。
绿原酸又名咖啡鞣酸(Chlorogenic acid，简称CHA)，是广泛存在于金银花、杜仲科植物杜仲的叶、葵花籽、茵陈和蓝盆花等植物中的一种有机酸，具有抗氧化、抗菌、抗病毒、降糖、降脂、降血压和免疫调节等多种药理作用。申请号为CN201510079639.8的中国专利申请公开了绿原酸在制备预防和治疗原发性皮肤T细胞淋巴癌的药物中的用途，该申请通过实验证明，绿原酸能够激活CD4T淋巴细胞和CD8T淋巴细胞，并以CD4T淋巴细胞和CD8T淋巴细胞为靶点，预防和治疗原发性皮肤T细胞淋巴癌。但是，原发性皮肤T细胞淋巴癌不是中枢神经系统肿瘤，它与中枢神经系统肿瘤的发病 原因、发病部位以及治疗机理大不相同。目前还未见将绿原酸用来治疗中枢神经系统肿瘤的报道。
发明内容
本发明的目的在于提供绿原酸在制备预防和/或治疗中枢神经系统肿瘤的药物中的新用途。
本发明提供了绿原酸在制备预防和/或治疗中枢神经系统肿瘤的药物中的用途。
进一步地，所述中枢神经系统肿瘤为中枢神经系统淋巴瘤。
进一步地，所述中枢神经系统淋巴瘤为原发性中枢神经系统淋巴瘤。
进一步地，所述原发性中枢神经系统淋巴瘤为B细胞非霍奇金淋巴瘤。
进一步地，所述B细胞非霍奇金淋巴瘤为弥漫性大B细胞淋巴瘤。
进一步地，所述弥漫性大B细胞淋巴瘤是脑部、眼部或脊髓部位的弥漫性大B细胞淋巴瘤。
进一步地，所述的药物是以绿原酸为活性成分，加上药学上可接受的辅料制备而成的药物制剂。
进一步地，所述的药物制剂为口服制剂。
进一步地，所述的药物制剂为注射制剂。
进一步地，所述的药物制剂的单位制剂中含绿原酸0.5～5.5mg。
进一步地，所述的药物制剂的单位制剂中含绿原酸1.1～3.3mg，优选3.3mg。
本发明还提供了一种治疗原发性中枢神经系统淋巴瘤的药物，它是以绿原酸为活性成分，加上药学上可接受的辅料制备而成的药物；所述药物的单位制剂中含绿原酸0.5～5.5mg，优选1.1～3.3mg，更优选3.3mg。
本发明首次发现绿原酸能够有效治疗弥漫性大B细胞淋巴瘤，为临床上中枢神经系统肿瘤，特别是弥漫性大B细胞淋巴瘤的治疗提供了一种新的、安全性高的选择，具有良好的应用前景。
显然，根据本发明的上述内容，按照本领域的普通技术知识和惯用手段，在不脱离本发明上述基本技术思想前提下，还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式，对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仅限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1为各给药组脑部弥漫性大B细胞淋巴瘤小鼠生长曲线情况。
图2为各给药组对小鼠的抑瘤率。
图3为各给药组眼眶弥漫性大B细胞淋巴瘤小鼠生长曲线情况。
图4为各给药组对小鼠的抑瘤率。
具体实施方式
本发明所用原料与设备均为已知产品，通过购买市售产品所得。
实施例1本发明绿原酸口服制剂处方
1、处方一
绿原酸1000g。
制备方法：按处方称取绿原酸，无菌分装成散剂。
2、处方二
绿原酸1000g、填充剂500g、粘合剂5g。
制备方法：按照处方称取绿原酸、填充剂、粘合剂，制粒，整粒、分装成颗粒剂。
3、处方三
绿原酸1000g、填充剂500g、粘合剂5g、润滑剂3g。
制备方法：按照处方称取绿原酸、填充剂、粘合剂，制粒，整粒，加润滑剂，压片，得片剂。
上述填充剂为甘露醇、乳糖、淀粉、微晶纤维素、糊精中的一种或多种；粘合剂为羧甲基纤维素钠、PVP(聚乙烯砒咯烷酮)中的一种或两种；润滑剂为硬脂酸镁、滑石粉、微粉硅胶中的一种或多种。
实施例2本发明绿原酸注射制剂处方
1、处方一
绿原酸1000g。
制备方法(1)：按处方无菌称取绿原酸，混合均匀后，无菌分装成粉针剂。
制备方法(2)：按照处方称取绿原酸，溶解于注射用水，过滤除菌，冷冻干燥，得冻干粉针剂。
2、处方二
绿原酸1000g、支架剂2667g、抗氧化剂67g。
制备方法：按照处方称取绿原酸、支架剂、抗氧化剂，溶解于注射用水，过滤除菌，冷冻干燥(冷冻干燥条件：预冻：温度≤-40℃，常压，干燥时间2-4h，冻结；一次干燥：温度≤-13℃，负压，干燥时间≥12h，充分干燥；二次干燥，温度20～30℃，负压，干燥时间≥2h)，得冻干粉针剂。
上述支架剂为甘露醇、乳糖、葡萄糖中的一种或多种；抗氧化剂为亚硫酸氢钠、维生素、谷胱甘肽、叶酸中的一种或多种。
以下通过实验例证明本发明的有益效果。
试验例1绿原酸治疗中枢神经系统淋巴瘤的体外试验
1、试验材料
1.1受试淋巴瘤细胞株
弥漫性大B细胞淋巴瘤细胞系pfeiffer购自美国ATCC，并以RPMI 1640培养基传代培养。
1.2受试药物
阳性药：注射用甲氨蝶呤，由江苏恒瑞医药股份有限公司提供。
受试药：绿原酸原料药。
以上两种药物均用无血清RPMI 1640培养基配制成所需浓度，后再经0.22μm微孔滤膜过滤除菌，放4℃冰箱备用。
1.3 RPML1640培养液
RPML1640细胞培养基溶解于超纯水(1000ml)中，搅拌溶解，加2.2g NaHCO 3和10ml HEPES搅拌溶解，再加青霉素适量(终浓度为100U/ml)和链霉素适量(终浓度为100μg/ml)，充分混合后，用0.22μm滤膜无菌过滤，分装，于-20℃冻存，此为基础培养基。临用前37℃水浴解冻，加10％的小牛血清，调整培养液pH为7.2～7.4，此为完全培养基。
2、试验方法
2.1细胞学实验
2.1.1细胞复苏
将装有细胞的冻存管投入37℃水浴中加热，快速晃动，尽快使其融化，融化后的细胞悬液放入10ml离心管，1000r/min低速离心5～10min，弃去上清后，加入适量新鲜培养基，轻轻吹打细胞，混匀后再次离心(1000r/min)5min，尽量减少培养基中的DMSO成分。再次离心后的细胞弃去上清，加新鲜培养基和10％的FBS，吹打细胞与培养基混合均匀，放入培养瓶，置于37℃恒温，5％CO 2的培养箱中培养。
2.1.2细胞培养方法
每日观察细胞培养基颜色及显微镜下观察细胞形态，并根据细胞状态，每隔1～2天更换培养基或传代，使细胞生长密度保持在4～6×10 5/ml。当培养基颜色由红变黄，需要换液时，将细胞置于50ml离心管中，1000r/min低速离心5～10min，弃上清，加入新鲜培养基后，轻轻吹打细胞，使细胞与培养基充分混匀，重新置于培养瓶内。细胞生长，密度增加，需要传代时，吸取一半体积的细胞液至另一培养瓶，分别加新鲜培养基至原体积。
2.1.3细胞冻存
取对数生长期的pfeiffer细胞，1000r/min低速离心5～10min，弃上清，细胞沉淀用冻存液重悬，进行细胞计数，仍用冻存液稀释，使细胞的密度在l×10 6～5×10 6/mL，将细胞液分装在冻存管内，标记细胞名称和冻存日期。依次放置在+4℃冰箱2h以上，-20℃冰箱24h以上，-70℃冰箱保存或再投入-196℃液氮罐中长期保存。
2.2 MTS法检测给药组对弥漫性大B细胞淋巴瘤细胞(pfeiffer)的体外抑瘤作用
2.2.1试验分组
实验分别分成8组：空白组、阴性对照组、阳性对照组及5组受试药物组。
空白组：只加入培养基，无需接种pfeiffer细胞；
阴性对照组：加培养基并接种pfeiffer细胞；
阳性对照组：10μg/ml的甲氨蝶呤与pfeiffer细胞进行培养；
受试药物组：受试药物为绿原酸，实验用浓度为5μg/ml、10μg/ml、30μg/ml、50μg/ml、100μg/ml，与pfeiffer细胞进行培养。
2.2.2细胞接种
取对数生长期细胞，0.25％胰蛋白酶消化，用基础培养基洗涤离心(1000r/min，5min两次)，用完全培养基悬浮细胞并调整细胞浓度为6×10 4/ml，接种细胞于96孔板内，每孔50μl(细胞数量6×10 3/孔)。
2.3加药
次日细胞贴壁后按分组加药，在37℃恒温，5％CO 2条件下静置培养48小时。每孔的加药剂量为100μl，空白组不接种细胞，加入100μl等量培养基，阴性对照组在接种细胞的基础上加入100μl等量培养基，每组3个复孔。
2.4染色
每孔加20μl MTS，继续培养3小时显色。检测前摇晃培养板10秒钟，混匀颜色。
2.5测定
使用酶联免疫检测仪测定OD值，用空白对照调零，所用波长为490nm。
2.6数据处理
按照以下公式计算肿瘤细胞的生长的抑制率：
GI(生长抑制率)＝(1-OD药物组/OD阴性对照组)×100％。
3、试验结果
3.1细胞形态观察结果
各组加药处理48小时后，在倒置显微镜下观察细胞形态，与阴性对照组比较，阳性对照组和绿原酸受试药物组对细胞生长有一定的影响，部分细胞变圆、脱落、悬浮；其中绿原酸受试药物组在50～100μg/ml的浓度下对pfeiffer癌细胞株生长抑制作用显著，大量细胞变圆、脱落、悬浮。
3.2各给药组对pfeiffer癌细胞的抑制率和剂量效应曲线
各给药组对pfeiffer癌细胞的抑制率见表1。
表1各给药组对pfeiffer癌细胞的抑制率
Figure PCTCN2022104224-appb-000001
从上表可见，绿原酸受试药物组对pfeiffer癌细胞的抑瘤作用明显，且在 绿原酸浓度为50～100μg/ml时，对肿瘤的抑制率显著，高达71.68～72.97％；此外，相同剂量(10μg/ml)的绿原酸对pfeiffer癌细胞的抑瘤作用高于阳性对照药物甲氨蝶呤。
上述实验结果表明，绿原酸对弥漫性大B细胞淋巴瘤细胞pfeiffer具有明显的体外抑制作用，在该体系下，绿原酸浓度为50～100μg/ml的抑瘤作用更为显著。
试验例2绿原酸治疗脑部弥漫性大B细胞淋巴瘤的动物体内试验
1、试验材料
1.1受试药物
受试药物1：注射用绿原酸，处方配比：绿原酸、甘露醇、亚硫酸氢钠(30:80:2)；
受试药物2：绿原酸口服制剂，处方配比：绿原酸、填充剂(乳糖)、粘合剂(羧甲基纤维素钠)(1000：500：5)；
阳性药物：甲氨蝶呤，江苏恒瑞医药股份有限公司。
以上注射用绿原酸制剂按处方配比称取绿原酸、甘露醇、亚硫酸氢钠，溶解于注射用水，过滤除菌，冷冻干燥(冷冻干燥条件：预冻：温度≤-40℃，常压，干燥时间2-4h，冻结；一次干燥：温度≤-13℃，负压，干燥时间≥12h，充分干燥；二次干燥，温度20～30℃，负压，干燥时间≥2h)，得绿原酸标示量为30mg/支的冻干粉针剂。
绿原酸口服制剂按处方配比称取绿原酸、乳糖、羧甲基纤维素钠混合，制粒，整粒，分装成颗粒剂。
1.2受试细胞株
弥漫性大B细胞淋巴瘤细胞Ly8购自复旦大学附属肿瘤医院，于10％小牛血清DMEM完全培养液中悬浮培养、传代，生长稳定。于细胞对数生长期时收集细胞，离心计数稀释细胞至10 4/μl浓度备用。接种细胞量2×10 4个/只。
1.3受试动物
BALB/c小鼠：60只，雌雄各半，鼠龄3～4周，体重15～22g，购自华西医学中心实验动物管理中心，饲养室内恒温、洁净并定期消毒，更换器具、垫料和饮水均在无菌条件下进行。
2、试验方法
2.1实验动物脑部弥漫性大B细胞淋巴瘤模型的建立
将小鼠经1％戊巴比妥钠0.2μl腹腔注射麻醉后，固定于立体定向仪，2％碘氯己锭溶液消毒头颅部皮肤。注射器进针部位为小鼠额部距颅中线左或右侧0.1cm，冠状缝前0.3cm处，将微量进样注射器依次穿透小鼠颅部皮肤和颅骨，进入颅内约1～2mm则固定注射器，缓慢注入Ly8细胞悬液2μl(约2×10 4个细胞)，在15min内注射完毕，然后留针5min使细胞充分沉积，再缓慢除针。术后无须注射抗生素。在麻醉后的睡眠中，小鼠保存在30℃2h。 待麻醉结束放置于25℃室温下，于接种24小时后观察表面变化。症状观察：注射侧头顶部隆起、身体消瘦、萎靡不振和活动度减低等。
2.2试验分组
将接种成功后的小鼠随机分成7组，每组6只，包括：
(1)阴性对照组：腹腔注射生理盐水，一天一次；
(2)阳性药物组：腹腔注射甲氨蝶呤，两天一次，3mg/kg/次；
(3)口服绿原酸颗粒组：口服受试药物2，一天一次，30mg/kg/次；
(4)注射给药组1：腹腔注射受试药物1，一天一次，5mg/kg/次；
(5)注射给药组2：腹腔注射受试药物1，一天一次，10mg/kg/次；
(6)注射给药组3：腹腔注射受试药物1，一天一次，30mg/kg/次；
(7)注射给药组4：腹腔注射受试药物1，一天一次，50mg/kg/次。
以上配制好的受试药物组，均按小鼠体重每10g腹腔注射0.2mL给药。每日给药前现用现配，给药14天。
2.3治疗作用评价
给药期间每隔一天观察并记录小鼠瘤体的生长情况、进食、活动状况等其他不良反应。给药14天后采用颈椎脱位法处死小鼠，按给药前编号分别测量并记录体重、肿瘤大小。根据肿瘤重量计算肿瘤抑制率(％)。体重及瘤重用均值±标准差
Figure PCTCN2022104224-appb-000002
表示，并进行各给药组与阴性对照组之间、各给药组与阳性对照甲氨蝶呤组之间的t检验。
Figure PCTCN2022104224-appb-000003
3、实验结果
3.1实验动物模型建立情况
60只BALB/c小鼠接种Ly8细胞建立移植瘤模型，初期接种部位可见一局限性隆起，于2-3天消失，5天左右时间部分BALB/c小鼠接种部位开始长出一小隆起；后该部位隆起逐渐生长，大约至14天左右时间，移植瘤最大直径长至5mm左右。共有45只BALB/c小鼠成功建立移植瘤模型并持续生长。接种成功率为75％。
3.2各实验组对脑部弥漫性大B细胞淋巴瘤的治疗作用
将接种成功后的小鼠选取42只，随机分为7组，按试验方案分别给药，记录小鼠的体重及瘤重变化，结果见表2及图1、图2。
表2各组小鼠治疗观察情况
Figure PCTCN2022104224-appb-000004
Figure PCTCN2022104224-appb-000005
注：*P<0.01，与阴性对照组比较； #P<0.05，与阳性对照组比较。
由表2和图1、图2可知：
(1)阳性药物组、30mg/kg口服绿原酸颗粒剂给药组及10～30mg/kg绿原酸注射剂给药剂量组对脑部弥漫性大B细胞淋巴瘤有良好的治疗效果，与阴性对照组有显著性差异(P<0.05)。
(2)阴性对照组有3只死亡，阳性对照组、口服颗粒组、5mg/kg及50mg/kg注射给药剂量组均各有1只死亡，10～30mg/kg注射给药组无小鼠死亡，说明注射用绿原酸有效剂量组的治疗效果良好；另外经密切观察，阴性对照组小鼠有皮毛缺少光泽、食欲不振、活动减少等表现，随着肿瘤负荷的增加，上述症状日益明显，有效剂量治疗组小鼠的症状有明显改善，未出现治疗用药相关不良反应。
(3)口服绿原酸颗粒剂组及绿原酸注射剂有效剂量组小鼠生长曲线呈逐步上升趋势，阴性对照组和阳性对照组呈下降趋势，5mg/kg及50mg/kg注射给药剂量组变化趋势不明显，说明绿原酸注射剂有效剂量组和口服绿原酸颗粒剂组对患病小鼠的生长发育有一定促进作用。
(4)10mg/kg～30mg/kg注射给药组与阳性对照组瘤重变化有显著性差异(P<0.05)，其中30mg/kg给药剂量下，注射用绿原酸对脑部弥漫性大B细胞淋巴瘤的治疗效果最佳，说明30mg/kg给药剂量为最优给药剂量。
上述实验结果表明，绿原酸对脑部弥漫性大B细胞淋巴瘤有良好的体内治疗效果，其中10mg/kg～30mg/kg的注射剂给药剂量抑瘤率较高，以30mg/kg给药剂量效果最佳。
根据徐叔云等人主编的《药理实验方法学》，按照单位重量的剂量来算，小鼠的剂量相当于人的9.1倍。因此，可以推论出绿原酸以3.3mg/kg的给药剂量注射到人体时对脑部弥漫性大B细胞淋巴瘤患者的治疗效果最佳。
试验例3绿原酸治疗眼眶弥漫性大B细胞淋巴瘤的动物体内试验
1、试验材料
1.1受试药物
受试药物1：注射用绿原酸，处方配比：绿原酸、甘露醇、亚硫酸氢钠(30:80:2)；
受试药物2：绿原酸口服制剂，处方配比：绿原酸、填充剂(乳糖)、粘合剂(羧甲基纤维素钠)(1000：500：5)；
阳性药物：甲氨蝶呤，江苏恒瑞医药股份有限公司。
以上注射用绿原酸制剂按处方配比称取绿原酸、甘露醇、亚硫酸氢钠，溶解于注射用水，过滤除菌，冷冻干燥(冷冻干燥条件：预冻：温度≤-40℃，常压，干燥时间2-4h，冻结；一次干燥：温度≤-13℃，负压，干燥时间≥12h，充分干燥；二次干燥，温度20～30℃，负压，干燥时间≥2h)，得绿原酸标示量为30mg/支的冻干粉针剂。
绿原酸口服制剂按处方配比称取绿原酸、乳糖、羧甲基纤维素钠混合，制粒，整粒，分装成颗粒剂。
1.2受试癌细胞株
弥漫性大B细胞淋巴瘤细胞系pfeiffer购自美国ATCC。
1.3受试动物
BALB/c小鼠：60只，雌雄各半，鼠龄3～4周，体重15～22g，购自华西医学中心实验动物管理中心，饲养室内恒温、洁净并定期消毒，更换器具、垫料和饮水均在无菌条件下进行。
2、试验方法
2.1 pfeiffer细胞准备
按照ATCC提供的方法37℃水浴复苏，RPMI-1640培养液培养，每隔1～2天更换培养基或传代。待细胞进入稳定增生状态时，选取2～3代活力良好的细胞备，用细胞计数板在显微镜下进行细胞计数，调整接种细胞浓度至1.5×10 8/ml，密封备用。
2.2实验动物眼眶弥漫性大B细胞淋巴瘤模型的建立
采用瘤细胞悬液接种法。在细胞接种前对所有小鼠进行铯137放射，对小鼠进行更进一步的免疫抑制，提高接种成功率，照射之后6小时内接种肿瘤细胞。眼周75％酒精消毒后，用l ml空针吸取0.1ml瘤细胞悬液，经眶周颞上方眼睑皮肤进针注射至眶内，拔针后按压5min，观察裸鼠注射后无明显异常，接种后，连续一周给予含庆大霉素10万U/L的高压灭菌水预防感染，每天给予5％葡萄糖口服加蛋黄喂养，增强营养。每天观察小鼠生长状态及局部肿瘤生长情况。
2.3试验分组
将接种成功后的小鼠选取42只，随机分成7组，每组6只，包括：
(1)阴性对照组：腹腔注射生理盐水，一天一次；
(2)阳性药物组：腹腔注射甲氨蝶呤，两天一次，3mg/kg/次；
(3)口服绿原酸颗粒组：口服受试药物2，一天一次，30mg/kg/次；
(4)注射给药组1：腹腔注射受试药物1，一天一次，5mg/kg/次；
(5)注射给药组2：腹腔注射受试药物1，一天一次，10mg/kg/次；
(6)注射给药组3：腹腔注射受试药物1，一天一次，30mg/kg/次；
(7)注射给药组4：腹腔注射受试药物1，一天一次，50mg/kg/次。
以上配制好的受试药物组，均按小鼠体重每10g腹腔注射0.2mL给药。每日给药前现用现配，给药14d。
2.3治疗作用评价
给药期间每隔一天观察并记录小鼠瘤体的生长情况、进食、活动状况等其他不良反应。给药14天后采用颈椎脱位法处死小鼠，按给药前编号分别测量并记录体重、肿瘤大小。根据肿瘤重量计算肿瘤抑制率(％)。体重及瘤重用均值±标准差
Figure PCTCN2022104224-appb-000006
表示，并进行各给药组与阴性对照组之间，各用药组与阳性对照甲氨蝶呤组和阴性对照组之间的t检验。
Figure PCTCN2022104224-appb-000007
3、实验结果
3.1实验动物模型建立情况
小鼠右眼眶内接种后可见眼球微凸，第2天恢复正常，第12天时，有40只BALB/C小鼠出现眼部症状，如眼睑睁不开，流泪，眼球周围球结膜轻度水肿，眼球无突出；第15天，55只BALB/C裸鼠出现眼部症状，流泪，眼球周围球结膜水肿明显，眼球无明显突出，接种成瘤率91.67％。
3.2各实验组对眼眶弥漫性大B细胞淋巴瘤的治疗作用
将接种成功后的小鼠选取42只，随机分为7组，按试验方案分别给药，记录小鼠的体重及瘤重变化，结果见表3及图3、图4。
表3各组小鼠治疗观察情况
Figure PCTCN2022104224-appb-000008
注：*P<0.01，与阴性对照组比较； #P<0.05，与阳性对照组比较。
由表3和图3、图4可知：
(1)阳性药物组、30mg/kg口服绿原酸颗粒剂给药组及10～30mg/kg绿原酸注射剂给药剂量组对眼眶弥漫性大B细胞淋巴瘤有良好的治疗效果，与阴性对照组有显著性差异(P<0.05)。
(2)阴性对照组有2只死亡，阳性对照组、口服颗粒组、5mg/kg及50mg/kg注射给药剂量组均各有1只死亡，10～30mg/kg注射给药组无小鼠死亡，说明注射用绿原酸有效剂量组的治疗效果良好；另外经密切观察，阴性对照组随着时间变化，眼球突出逐渐明显，肿瘤逐渐包裹眼球，最后致眼睑无法闭合，眼球被遮挡，肿物暴露在外破溃出血，流泪增多。口服绿原酸 颗粒组、10～30mg/kg注射给药组小鼠眼睛不适能得到很大的改善，脓肿减少，流泪症状缓解。
(3)口服绿原酸颗粒剂组及绿原酸注射剂有效剂量组小鼠生长曲线呈逐步上升趋势，阴性对照组和阳性对照组呈下降趋势，5mg/kg及50mg/kg注射给药剂量组变化趋势不明显，说明绿原酸注射剂有效剂量组和口服绿原酸颗粒剂组对患病小鼠的生长发育有一定促进作用。
(4)10mg/kg～30mg/kg注射给药组与阳性对照组瘤重变化有显著性差异(P<0.05)，其中30mg/kg给药剂量下，注射用绿原酸对眼眶弥漫性大B细胞淋巴瘤的治疗效果最佳，说明30mg/kg给药剂量为最优给药剂量。
上述实验结果表明，绿原酸对眼眶弥漫性大B细胞淋巴瘤有良好的体内治疗效果，其中10mg/kg～30mg/kg的注射剂给药剂量抑瘤率较高，以30mg/kg给药剂量效果最佳。可以推论出绿原酸以3.3mg/kg的给药剂量注射到人体时对眼眶弥漫性大B细胞淋巴瘤患者的治疗效果最佳。
综上，本发明提供了绿原酸在制备预防和/或治疗中枢神经系统肿瘤的药物中的用途。本发明首次发现绿原酸能够有效治疗弥漫性大B细胞淋巴瘤，为临床上中枢神经系统肿瘤，特别是弥漫性大B细胞淋巴瘤的治疗提供了一种新的、安全性高的选择，具有良好的应用前景。

Claims (10)

  1. 绿原酸在制备预防和/或治疗中枢神经系统肿瘤的药物中的用途。
  2. 根据权利要求1所述的用途，其特征在于：所述中枢神经系统肿瘤为中枢神经系统淋巴瘤。
  3. 根据权利要求2所述的用途，其特征在于：所述中枢神经系统淋巴瘤为原发性中枢神经系统淋巴瘤。
  4. 根据权利要求3所述的用途，其特征在于：所述原发性中枢神经系统淋巴瘤为B细胞非霍奇金淋巴瘤。
  5. 根据权利要求4所述的用途，其特征在于：所述B细胞非霍奇金淋巴瘤为弥漫性大B细胞淋巴瘤。
  6. 根据权利要求5所述的用途，其特征在于：所述弥漫性大B细胞淋巴瘤是脑部、眼部或脊髓部位的弥漫性大B细胞淋巴瘤。
  7. 根据权利要求1～6任一项所述的用途，其特征在于：所述药物是以绿原酸为活性成分，加上药学上可接受的辅料制备而成的药物制剂。
  8. 根据权利要求7所述的用途，其特征在于：所述的药物制剂为口服制剂或注射制剂。
  9. 根据权利要求8所述的用途，其特征在于：所述的药物制剂的单位制剂中含绿原酸0.5～5.5mg，优选1.1～3.3mg，更优选3.3mg。
  10. 一种治疗中枢神经系统肿瘤的药物，它是以绿原酸为活性成分，加上药学上可接受的辅料制备而成的药物；所述药物的单位制剂中含绿原酸0.5～5.5mg，优选1.1～3.3mg，更优选3.3mg。
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