WO2007036113A1 - Usage médical du periplocoside a et e - Google Patents

Usage médical du periplocoside a et e Download PDF

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WO2007036113A1
WO2007036113A1 PCT/CN2006/001783 CN2006001783W WO2007036113A1 WO 2007036113 A1 WO2007036113 A1 WO 2007036113A1 CN 2006001783 W CN2006001783 W CN 2006001783W WO 2007036113 A1 WO2007036113 A1 WO 2007036113A1
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preparation
immunosuppressive
disease
medicament
treating
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Weimin Zhao
Jianping Zuo
Qunfang Liu
Yina Zhu
Jian Zou
Yifu Yang
Jia Tian
Yu Zhou
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Shanghai Institute Of Materia Medica, Chinese Academy Of Sciences
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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
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    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
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    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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Definitions

  • the present invention relates to the pharmaceutical use of the natural products of periplocin A and E, and in particular to the use of the natural products of luciferin A and E for the preparation of immunosuppressive drugs. Background technique
  • Immunosuppressors are drugs that inhibit immunity, and different specific drugs are used as immunosuppressants for different aspects of immune response and regulation. It is mainly used clinically to treat autoimmune diseases and prevent organ transplant rejection. Immunosuppressive agents are mainly classified into the following types: (1) Chemical preparations, such as nitrogen mustards of alkylating agents. (2) Hormones such as glucocorticoids. (3) Fungal metabolites, such as cyclosporin A (CsA (4) Chinese medicine and its active ingredients.
  • CsA cyclosporin A
  • immunosuppressive drugs have obvious toxic side effects, mainly myelosuppression, liver and kidney toxicity. Since the action of immunosuppressive drugs is non-specific, it can lead to a decrease in the body's immune function, an increase in pathogenic microbial infections, and a long-term application may increase the incidence of cancer. Since most immunosuppressive agents have certain toxic side effects, it is important to find new high-efficiency and low-toxic immunosuppressive agents.
  • Xiangjiapi is the root bark of Periploca sepium Bge. Also known as goat milk, pilose peach, sheep cross. The main production is Shanxi, Henan, Hebei, Shandong, Gansu, Hunan and other provinces. Xiangjiapi has been used as a medicine for five years, and the product is called Xiangjiapi or Beijijia.
  • the compound periplocin A is a progestin compound (Periplocoside E) isolated from the bark, and its chemical name is: A 5 -pregnene-3 , 17 ⁇ , 20(S)-triol 3-0- ( 4', 6 '-dideoxy-3 ' -O-methyl- ⁇ 3 -D-2'- exosuloside ) 20-O-(2-O-acetyl- -D-digitalopyranosyl (1->4)- ⁇ -D -cymaropyranosyl (1->4)- -D-cymaropyranosy( 1 - ⁇ -4)- -D-cymaropyranosyl(l - 5)-3 ,7-dideoxy-4-0-methyl-aD-gluco-2-h
  • Eptulopyranosyl(2-4)-dioxy-(l ⁇ 3)-pD-canaropyranoside) is a natural compound with the molecular formula: C 72 H 114
  • the compound periplocin E is a progestin compound (Periplocoside E) isolated from the bark, and its chemical name is:
  • Eptulopyranosyl(2-4)-dioxy-(l - 3)-pD-canaropyranoside) is a natural compound whose molecular formula is:
  • the object of the present invention is to provide a pharmaceutical use of the natural products of periplocin A and E, which can be used for the preparation of immunosuppressive drugs.
  • saponin A and E can be used to prepare immunosuppressive drugs, such as liquid dosage forms such as injections, solutions, or oral dosage forms such as tablets and capsules under sterile conditions.
  • immunosuppressive drugs such as liquid dosage forms such as injections, solutions, or oral dosage forms such as tablets and capsules under sterile conditions.
  • Figure 1 shows the inhibitory effect of periplocin on the proliferation of ConA-stimulated T lymphocytes.
  • Figure 2 is a graph showing the inhibitory effect of periplocin E on LPS-stimulated lymphocyte proliferation.
  • Figure 3 is the toxicity of periplocin E to lymphocytes.
  • Figure 4 is a graph showing the inhibitory effect of periplocin on the proliferation of allogeneic mixed lymphocytes.
  • Figure 5 is a graph showing the inhibitory effect of periplocin E on delayed hypersensitivity in mice. detailed description
  • Test animals Pure Balb/c female mice, 6-8 weeks old.
  • RPMI-1640 medium Gibco, pH 7.2 was supplemented with 10% fetal bovine serum (FBS), 100' U/ml penicillin, 100 ⁇ g/ml streptomycin, 10 mM HEPES, and 50 ⁇ 2- ⁇ .
  • mice were sacrificed from the spine, the spleen was aseptically taken, ground to make a single cell suspension, and the MTT solution (10% SDS, 50% dimethylformamide; PH7.2) was used to remove the red blood cells.
  • the %FBS RPMI-1640 medium was adjusted to a cell concentration of 5 x 107 ml.
  • mice were sacrificed by depolarization, and the spleen was aseptically prepared to prepare a single cell suspension, and the cell concentration was adjusted to 4 x 107 ml with RPMI-1640 medium containing 10% FBS.
  • mice were sacrificed by defibrillation, the spleen was aseptically taken, ground to make a single cell suspension, and after removing red blood cells, the cell concentration was adjusted with RPMI-1640 medium containing 10% calf serum. Adjust to 6xl07ml.
  • C57BL/6 spleen cells are reactive cells
  • Balb/c spleen cells are stimulator cells
  • the two cells are mixed in equal volumes.
  • 3. Add 100 ⁇ l of C57BL/6 and Balb/c splenocyte mixture to a 96-well plate, and sample 100 ⁇ l. Add 100 ⁇ l of culture medium containing 10% serum. And do a separate culture control of the two cells.
  • the plate is frozen in a _20 °C refrigerator.
  • the cell harvester (HARVESTER96®, T0MTEC) collects the cells on the glass fiber membrane and uses a liquid flash counter (MicroBeta Trilux®, PerkinElmer) to detect the incorporation of DNA into [ ]-thymidine, reflecting cell proliferation. happening.
  • the effect of the immune response process is mainly composed of a T cell-mediated cellular immune response and a B cell-mediated humoral immune response.
  • periplocin E and periplocin A showed good inhibitory activity against lymphocyte proliferation.
  • the inhibition rate of conjugated T cell proliferation was 72%, and the inhibition rate of LPS-stimulated B cell proliferation was 53%;
  • the inhibition rate of T cell proliferation was 47%, and the inhibition rate of LPS-stimulated B cell proliferation was 26%. Therefore, we chose a relatively more active oleosin E for further study.
  • the TC50 of periplocin E is 10. 1 ⁇ 1. 4 ⁇ M, and there is no lymphocyte toxicity at less than 5 ⁇ M.
  • Inhibition of ConA-stimulated T cell proliferation IC50 is 0. 64 +0. 23 ⁇ ⁇ ; inhibition of LPS-stimulated B cell proliferation IC50 is 0.90 ⁇ 0.17 ⁇ ⁇ .
  • Allogeneic antigens are the main cause of body rejection after transfusion and organ transplantation.
  • the allogeneic antigens expressed on the heterologous lymphocytes mainly the histocompatibility antigens MHC-I and MHC-II, stimulate the T cells to elicit an immune proliferative response.
  • MHC-I and MHC-II the histocompatibility antigens
  • MHC-I and MHC-II mainly the histocompatibility antigens
  • the serotonin E significantly inhibited the proliferation of responsive lymphocytes in the MLR with an EC50 of 0.25 ⁇ 0.13 ⁇ .
  • the DTH assay is a method of detecting cell-mediated immune function in vivo, similar to contact hypersensitivity.
  • DTH also includes two stages of sensitization and triggering. Sensitization takes about 6 days, and the inflammatory response of DTH peaks 24 to 48 hours after the trigger. In the initial sensitization phase, memory-specific antigen-specific cells are distributed to peripheral lymphoid tissues, and any localized antigenic stimulation will rapidly activate these cells to cause DTH.
  • periplocin E at 5 mg/kg and 20 mg/kg significantly inhibited ear swelling (P ⁇ 0.01).

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Description

杠柳苷 A和 E的医学用途 技术领域
本发明涉及天然产物杠柳苷 A和 E的药物用途， 具体地说， 本发明是关于天然产物杠 柳苷 A和 E用于制备免疫抑制药物的用途。 背景技术
免疫抑制剂 （immunosuppressor) 是对免疫有抑制作用的药物， 不同的具体药物分别作 用于免疫反应及调节的不同环节免疫抑制剂。在临床上主要用于治疗自身免疫性疾病和防止 脏器移植排斥。 免疫抑制剂主要分为以下几种： （1 )化学制剂类， 如烷化剂类的氮芥。 （2) 激素类， 如糖皮质激素。 （3 )真菌代谢产物， 如环孢素 A (cyclosporinA,CsA (4) 中药及 其有效成分。
' 免疫抑制药物大多具有明显的毒副作用， 主要是骨髓抑制， 肝、 肾毒性等。 由于免疫抑 制药物的作用是非特异的， 所以可导致机体免疫功能的下降， 病原微生物感染增加， 长期应 用可能提高肿瘤发病率。 由于免疫抑制剂大多数具有一定的毒副作用， 因此， 寻找新的高效 低毒的免疫抑制剂具有重要的意义。
香加皮为萝蘼科植物（Periploca sepium Bge. )杠柳的根皮。 又名羊奶条， 羊角桃， 羊交 叶。主产山西、河南、河北、 山东、甘肃、湖南等省。香加皮多年来一直被当作五加皮入药， 商品习称香加皮或者北五加。
化合物杠柳苷 A为从杠柳中分离的到的一种孕烷苷类化合物 （Periplocoside E) ，化学名 为： A5-pregnene-3 , 17α, 20(S)-triol 3-0- (4', 6 ' -dideoxy-3 ' -O-methyl-Δ3 -D-2'- exosuloside ) 20-O-(2-O-acetyl- -D-digitalopyranosyl (1->4)- β-D-cymaropyranosyl (1->4)- -D-cymaropyranosy( 1 -→-4)- -D-cymaropyranosyl(l - 5)-3 ,7-dideoxy-4-0-methyl-a-D-gluco-2-h eptulopyranosyl(2-4)-dioxy-(l→3)-p-D-canaropyranoside)，是一种天然化合物， 其分子式为： C72H114027, 据文献报导， 杠柳苷 A具有一定的抗肿瘤活性。
其化学结构如下：
Figure imgf000004_0001
化合物杠柳苷 E为从杠柳中分离的到的一种孕烷苷类化合物 （Periplocoside E) ，化学名 为 ： A5-pregnene-3p, 17α, 20(S)-triol 20-(2-O-acetyl-p-D-digitalopyranosyl (1→ )- β-D-cymaropyranosyl (1- 4)- -D-cymaropyranosy(l->4)-P-D-cymaropyranosyl(l→5)-3,7-dideoxy-4-0-methyl-a-D-gluco-2-h eptulopyranosyl(2-4)-dioxy-(l - 3)-p-D-canaropyranoside) , 是一种天然化合物， 其分子式为：
C65H106O24, 其化学结构如下：
Figure imgf000005_0001
目前尚未发现有天然产物杠柳苷 A, E具有免疫抑制活性的报道。
发明内容
本发明的目的就在于提供天然产物杠柳苷 A和 E的药物用途， 它们可以用于制备免疫 抑制药物。
本申请的发明人通过研究发现， 天然产物杠柳苷 A和 E具有免疫抑制作用。 因此， 杠 柳苷 A和 E可以用來制备免疫抑制药物， 如在无菌条件下制成液体剂型如针剂、 溶液， 或 者是口服剂型如片剂和胶囊等。 附图说明
图 1是杠柳苷 E对 ConA刺激的 T淋巴细胞增殖的抑制作用。
图 2是杠柳苷 E对 LPS刺激的淋巴细胞增殖的抑制作用。
图 3是杠柳苷 E对淋巴细胞的毒性。
图 4是杠柳苷 E对同种异体混合淋巴细胞增殖反映的抑制作用。
图 5是杠柳苷 E对小鼠迟发性超敏反应的抑制作用。 具体实施方式
以下通过具体实施例来进一步说明本发明。应理解， 以下实施例仅用于说明本发明而不 用于限定本发明的范围。
实施例 1 北五加中活性成分杠柳苷 E的免疫抑制作用
一. 实验材料
试验动物： 纯系 Balb/c雌性小鼠， 6- 8周龄。
RPMI-1640 培养液 (Gibco, pH7. 2 ) 添加 10%胎牛血清 (FBS ) ， 100 ' U/ml 青霉素， 100 μ g/ml 链霉素， lOmM HEPES, 及 50 μ Μ 2-ΜΕ。 刺激剂： 伴刀豆蛋白 （ConA)， 细菌脂多糖 （LPS, 来自 £sc ?eric 's (¾/ 055 : B5)， 临 用前以 RPMI- 1640培养液稀释至合适浓度。 二. 实验方法
[一] 淋巴细胞毒性实验
1.脊椎处死小鼠， 无菌取其脾脏， 磨碎制成单细胞悬液， 用 MTT溶解液 （10%SDS， 50 %二甲基甲酰胺； PH7. 2) 去除红细胞后， 用含 10%FBS 的 RPMI- 1640 培养液将细胞浓度调 成 5xl07ml。
2. 96孔培养板中加入 80 μ ΐ的细胞悬液， 40 μ 1的样品， （补充样品的浓度， 是什么样 品？ 是 Α还是 E? ) 40 μ 1含 10%FBS的培养液； 对照加 80 μ 1培养液， 总体积为 160 μ 1。 并设空白对照。
3.放入 37° C, 5% C02培养箱中培养 48小时。在结束培养前 6-7小时，每孔加 5mg/ml MTT 16 μ 1。
4.结束培养吋， 每孔加 80 μ 1 MTT溶解液 ( 10%SDS, 50%二甲基甲酰胺; pH7. 2)， 在 培养箱中放置 6-7小时后， 用酶标仪于 570nm处测定 0D57。值。
[二] 淋巴细胞增殖实验
1.脱脊椎处死小鼠， 无菌取脾脏制成单细胞悬液， 用含 10%FBS的 RPMI- 1640 培养液将 细胞浓度调成 4xl07ml。
2.于 96孔板中加入 100 μ ΐ的细胞悬液， 50 μ 1的样品溶液， 50 μ 1 ConA或 LPS溶液， 对照孔加 50 μ 1含 10%FBS的培养液。 总体积为 200 μ 1。
3.于 37° C, 5% C02培养箱中培养 48小时。在结束培养前 7-8小时，每孔加 0. 5 μ Ci [¾]- 胸腺嘧啶核苷 （25 μ 1/孔）。
4.结束培养， 用细胞收集仪 （HARVESTER96®，T0MTEC) 收集细胞于玻璃纤维膜上， 加入 闪烁液， 用液体闪烁记数仪 （MicroBeta Trilux®, PerkinElmer) 检测细胞 DNA 中 [¾] -胸 腺嘧啶核苷的掺入量， 反映细胞增殖情况。
[三] 同种异体混合淋巴细胞增殖反应 (MLR)
1.脱脊椎处死 C57BL/6和 Balb/c小鼠， 无菌取其脾脏， 磨碎制成单细胞悬液， 去除红 细胞后， 用含 10%小牛血清的 RPMI- 1640 培养液将细胞浓度调成 6xl07ml。
2. C57BL/6脾细胞为反应细胞， Balb/c脾细胞（经钴 60照射， 3000 rads )为剌激细胞， 两种细胞等体积混合。 3.于 96孔板中加入 C57BL/6和 Balb/c脾细胞混合液 100 μ 1 , 样品 100 μ 1， 对照加 100 μ 1含 10%血清的培养液。 并做两种细胞的单独培养对照。
4. 37° C， 5% CO2培养箱中培养 3， 4, 5天。 在结束培养前 1天， 加入 ¾稀释液 25 ΐ (即， 3. 8xl0' q的 [ ]-胸腺嘧啶核苷）。
5.结束培养时， 将培养板冻存于 _20°C冰箱。
6.细胞收集仪（HARVESTER96® , T0MTEC)收集细胞于玻璃纤维膜上， 用液闪记数仪 (MicroBeta Trilux®, PerkinElmer) 检测 DNA对 [ ]-胸腺嘧啶核苷的掺入量， 反映细胞 增殖情况。
[四] 迟发性超敏反应动物模型 （DTH)
1. DayO, Balb/c小鼠 （雌性， 6-8周龄） 每组八只， 每只后脚以 20 μ 1 0. 5% DNFB致 敏。 次 R加强。 （DNFB溶于 [丙酮： 橄榄油 =4: 1]油剂中）；
2. Day ( 7-9)， 鼠右耳内外两侧各 10 μ 1 0. 4% DNFB进行攻击；
3.攻击前 1天腹腔注射一次， 攻击前腹腔注射一次， 24小时后， 再给药一次；
4.攻击后 30- 48小时， 检测各指标。 三. 实验结果
[一] 五加皮提取物对淋巴细胞增殖的抑制活性及其毒性
免疫应答过程的效应表现主要是由以 T细胞介导的细胞免疫反应和 B细胞介导的体液 免疫反应组成。 我们在体外用 T细胞丝裂原 ConA和 B细胞丝裂原 LPS直接刺激细胞和细胞 的活化分裂增殖来评价药物淋巴细胞增殖反应的抑制活性。在诸多五加皮提取物中， 杠柳苷 E和杠柳苷 A表现出对淋巴细胞增殖有较好的抑制活性。 在无淋巴细胞毒剂量 ΙμΜ 时， 杠 柳苷 Ε对 ConA刺激的 T细胞增殖的抑制率是 72%， 对 LPS刺激的 B细胞增殖的抑制率是 53% ；杠柳苷 A对 ConA剌激的 T细胞增殖的抑制率是 47%，对 LPS剌激的 B细胞增殖的抑 制率是 26% 所以我们选择活性相对更强的杠柳苷 E作经一步研究。 表： 五加皮活性提取物对淋巴细胞增殖的抑制活性及其毒性 化合物 浓度 Con A刺激的 LPS剌激的 淋巴细胞
(μΜ) T淋巴细胞增殖的 B淋巴细胞增殖的 毒性
抑制率 抑制率
杠柳苷 E 10 83% 76% 50%
1 72% 53% 无毒
0.1 26% 23% 无毒
杠柳苷 A 10 84% 36 % 42%
1 47% 26% 无毒
0.1 21% 13% 工
尤母 [二] 杠柳苷 E对淋巴细胞增殖的作用和毒性
杠柳苷 E的 TC50是 10. 1 ± 1. 4 μ M, 在低于 5 μ Μ时没有淋巴细胞毒性。 对 ConA刺激的 T细胞增殖的抑制 IC50是 0. 64 +0. 23 μ Μ; 对 LPS刺激的 B细胞增殖的抑制 IC50是 0. 90 ±0. 17 μ Μ。 综合以上结果， 我们在体内和体外进一步检査其免疫抑制药理活性。
[二] 杠柳苷 Ε对同种异体混合淋巴细胞增殖反应 (MLR)的作用
异体抗原是输血、 器官移植后引起机体排异反应的主要原因。 应答淋巴细胞和异源性淋 巴细胞共培养时， 表达在异源性淋巴细胞上的异体抗原， 主要是组织相容性抗原 MHC-I、 MHC-II分子， 刺激应答 T细胞引发免疫增殖反应。 我们通过药物对 MLR的作用来评价药物 对更接近生理的免疫系统应答的药理作用。杠柳苷 E显著的抑制了 MLR中应答淋巴细胞的增 殖， 其 EC50是 0. 25±0. 13 μ Μ。
[三] 杠柳苷 Ε对迟发性超敏反应动物模型 （DTH) 的作用
为了进一步检査在体内的免疫药理活性。 我们选用了经典的小鼠 DTH反应模型。 DTH 实验是一种体内捡测细胞介导免疫功能的方法， 与接触性超敏反应相似。 DTH也包括致敏和 触发两个阶段。 致敏约需要 6天， 在触发后的 24〜48小时 DTH的炎症反应达到高峰。 在初 始的致敏阶段有记忆性的抗原特异性细胞分布到外周各淋巴组织，在任何局部的抗原剌激会' 使这些细胞迅速活化引起 DTH。 结果显示， 体内给药杠柳苷 E在 5mg/kg, 20mg/kg时， 能显 著抑制耳肿胀程度 （P〈0. 01 )。 与对照组相比抑制率分别 53. 9%和 66. 6%。 小结： 天然产物杠柳苷 A和 E对于小鼠的 T淋巴细胞与 B淋巴细胞增殖具有明显的抑 制作用， 在体内实验中， 杠柳苷 A和 E对于 DTH反应模型的小鼠的耳肿胀程度都具有显著 的抑制作用 （P<0. 01 )， 且毒性较低。 可以进一步将其开发成为高效低毒的免疫抑制药物， 如在无菌条件下被制成液体剂型如针剂、 溶液， 或者是口服剂型如片剂和胶囊等。 由于这些 技术都是本领域的常规技术手段， 因此不再赘述。

Claims

1.结构式如下的天然产物杠柳苷 A和 E用于制备免疫抑制剂， 包括作为活性成分的 T、 Β淋巴细胞的增殖反应抑制剂中应用。
权
求
Figure imgf000009_0001
(2)杠柳苷 Ε
2.根据权利要求 1所述用于制备免疫抑制药物中应用，其特征在于制备治疗自身免疫疾 病的药物中应用。
3.根据权利要求 2所述用于制备免疫抑制药物中应用，其特征在于免疫性疾病为类风湿 性关节炎、 自身免疫性肝炎、 胰岛素依赖性糖尿病、 溃疡性结肠炎、 多发性硬化、 硬皮病、 重症肌无力、 多发肌炎、 自身免疫性血细胞减少症、 脉管炎综合症或系统性红斑狼疮。
4.根据权利要求 1 所述用于制备免疫抑制药物中应用， 其特征在于在制备治疗炎症反 应、 纤维化或由自身免役疾病及相关疾病所致具有组织损伤或感染的功能异常药物中应用。
5.根据权利要求 1所述用于制备免疫抑制药物中应用，其特征在于在制备治疗过敏性疾 病药物中应用。
6.根据权利要求 5所述用于制备免疫抑制药物中应用，其特征在于过敏性疾病为支气管 哮喘、 过敏性鼻炎、 特应性皮炎或花粉病。
7.根据权利要求 1所述用于制备免疫抑制药物中应用，其特征在于在制备治疗排异反应 的药物中应用。
8.根据权利要求 1所述用于制备免疫抑制药物中应用，其特征在于在制备治疗由骨髓造 血干细胞移植所致的移植物抗宿主疾病药物中应用。
9.权利 1到 8项中任一项的免疫抑制剂，其中所述的抑制剂包括具有权利要求 9所列结 构式的化学成分作为活性成分的植物提取物及复方药物 。
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108785317A (zh) * 2017-04-28 2018-11-13 苏州凯祥生物科技有限公司 香加皮c21甾类化合物在制备ido抑制剂中的用途
CN108785322A (zh) * 2017-04-28 2018-11-13 南京葆赫生物技术有限公司 黑骨藤c21甾类在制备ido抑制剂中的用途

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102532229B (zh) * 2010-12-15 2015-01-07 中国科学院上海药物研究所 一类含有原酸酯基团的孕甾烷糖苷类化合物及其用途
CN102675394B (zh) * 2011-03-08 2015-09-30 西北农林科技大学农药研究所 一种杠柳新苷类农用杀虫化合物
CN103130903A (zh) * 2011-11-23 2013-06-05 杨凌农科大农药研究所 杠柳新苷p-一种对昆虫胰蛋白酶具有激活作用的农用杀虫活性化合物
CN108785316B (zh) * 2017-04-28 2021-02-05 苏州凯祥生物科技有限公司 香加皮c21甾类在制备ido抑制剂中的用途
CN108785321B (zh) * 2017-04-28 2021-02-05 南京葆赫生物技术有限公司 黑骨藤c21甾类化合物在制备ido抑制剂中的用途
CN113577085B (zh) * 2020-04-30 2023-04-18 中国科学院上海药物研究所 杠柳苷组合物、其制备方法及其在制备治疗自身免疫性疾病的药物中的用途

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1569088A (zh) * 2003-07-11 2005-01-26 中国人民解放军军事医学科学院毒物药物研究所 黑骨藤提取物，其药物组合物及其药物用途

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ITOKAWA H.: "Studies on Chemical Constituents of Antitumor Fraction from Periploca sepium. II. Structures of New Pregnance Glycosides, Periplocosides A, B and C", CHEMICAL & PHARMACEUTICAL BULLETIN, vol. 36, no. 3, March 1988 (1988-03-01), pages 982 - 987, XP003011362 *
ITOKAWA H.: "Studies on Chemical Constituents of Antitumor Fraction from Periploca sepium. IV. Structures of New Pregnance Glycosides. Periplocosides A, B and C", CHEMICAL & PHARMACEUTICAL BULLETIN, vol. 36, no. 3, March 1988 (1988-03-01), pages 982 - 987, XP008079669 *
LI H. AND LI G.: "Recent Advances of Periploca Plants", CHINESE TRADITIONAL AND HERBAL DRUGS (CHINESE), vol. 23, no. 3, March 1992 (1992-03-01), pages 150 - 158, XP008079668 *
ZHANG Y. AND WANG F.: "Recent Advances of Chemical Constituents of Periploca Plants", NATURAL PRODUCT RESEARCH AND DEVELOPMENT (CHINESE), vol. 15, no. 2, 2003, pages 157 - 161, XP008079667 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108785317A (zh) * 2017-04-28 2018-11-13 苏州凯祥生物科技有限公司 香加皮c21甾类化合物在制备ido抑制剂中的用途
CN108785322A (zh) * 2017-04-28 2018-11-13 南京葆赫生物技术有限公司 黑骨藤c21甾类在制备ido抑制剂中的用途
CN108785322B (zh) * 2017-04-28 2020-11-27 南京葆赫生物技术有限公司 黑骨藤c21甾类在制备ido抑制剂中的用途
CN108785317B (zh) * 2017-04-28 2020-12-01 苏州凯祥生物科技有限公司 香加皮c21甾类化合物在制备ido抑制剂中的用途

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