WO2021218965A1 - 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用 - Google Patents

茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用 Download PDF

Info

Publication number
WO2021218965A1
WO2021218965A1 PCT/CN2021/090253 CN2021090253W WO2021218965A1 WO 2021218965 A1 WO2021218965 A1 WO 2021218965A1 CN 2021090253 W CN2021090253 W CN 2021090253W WO 2021218965 A1 WO2021218965 A1 WO 2021218965A1
Authority
WO
WIPO (PCT)
Prior art keywords
acid
cyclic peptide
cancer
cgas
type cyclic
Prior art date
Application number
PCT/CN2021/090253
Other languages
English (en)
French (fr)
Inventor
谭宁华
王琛
汪哲
王佳
Original Assignee
中国药科大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 中国药科大学 filed Critical 中国药科大学
Publication of WO2021218965A1 publication Critical patent/WO2021218965A1/zh

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/12—Cyclic peptides, e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia

Definitions

  • the invention belongs to pharmaceutical technology, and specifically relates to the application of a class of Rubiaceae type cyclic peptides in the preparation of drugs for cGAS-STING signal pathway activators.
  • Innate immunity is the host’s first line of defense against the invasion of pathogenic microorganisms.
  • the pattern recognition receptor of host cells can recognize the conservative components of pathogenic microorganisms and pathogen-related molecular patterns, such as nucleic acid molecules, and sense the invasion of pathogenic microorganisms through related signal transduction levels.
  • the joint reaction finally induces the expression of type I interferon and inflammatory factors, and finally eliminates the invading pathogenic microorganisms.
  • the recognition of "non-self" RNA and the signal transduction mechanism after recognition have been analyzed in detail, while the research on the recognition signal transduction mechanism of "non-self" DNA has only begun in recent years.
  • the cGAS-STING signal pathway is a key signal pathway for DNA recognition in cells. It plays an important role in the production of interferon and related inflammatory factors. It helps the body recognize and eliminate invading DNA pathogenic microorganisms. Under the stimulation of "non-self" DNA, cGAS quickly recognizes DNA, uses ATP and GTP to synthesize cGAMP, which is recognized by the node molecule STING on the endoplasmic reticulum, and then rapidly dimerizes and transfers from the endoplasmic reticulum to the perinuclear body At the same time, TBK1 is recruited and transferred to the perinuclear body to activate, and then phosphorylates the transcription factor IRF3; activated IRF3 dimerizes and merges into the nucleus, activates downstream interferon and related target gene expression, and participates in antiviral, Various biological effects such as inflammation and immune response.
  • This pathway is also closely related to the occurrence and development of many diseases, such as tumors, Aicardi-Goutieres syndrome, and systemic lupus erythematosus or lupus-like diseases. Therefore, in drug development, the discovery of modulators of small molecule compounds in the cGAS-STING signaling pathway, including activators or inhibitors, is a very promising research direction.
  • This application discloses the application of a Rubiaceae type cyclic peptide compound or a pharmacologically acceptable salt thereof in the preparation of drugs for cGAS-STING signal pathway activator.
  • the Rubiaceae type cyclic peptide compound is a kind of bicyclic ring hexapeptide compound, which is substituted by one D-type ⁇ -alanine, one L-type ⁇ -alanine, and three L-type N.
  • -Tyrosine and one other L-encoded ⁇ -amino acid are connected by a peptide chain to form a cyclic hexapeptide, and 6 amino acids are condensed into an eighteen-membered ring.
  • the Rubiaceae type cyclic peptide compound is the Rubiaceae type cyclic peptide 1-27 represented by the following structural formula:
  • the Rubiaceae type cyclic peptide compound described in this application is extracted from the roots and rhizomes of Rubia plants.
  • the above-mentioned drugs can be used to prevent and treat diseases related to the cGAS-STING signaling pathway, and the diseases are cancers, including colon cancer, rectal cancer, basal cell carcinoma, medulloblastoma, rhabdomyosarcoma, and nevus-like basal cell carcinoma syndrome.
  • Small cell lung cancer non-small cell lung cancer, metastatic prostate cancer, pancreatic cancer, chondrosarcoma, osteosarcoma, melanoma, glioma, breast cancer, ovarian cancer, esophageal cancer, gastric cancer, cholangiocarcinoma, liver cancer, bladder cancer , Hemangioma, chronic myeloid leukemia, acute lymphocytic leukemia, multiple myeloma, Hodgkin's lymphoma, non-Hodgkin's lymphoma.
  • the Rubiaceae type cyclic peptide compound or its pharmacologically acceptable salt can activate the expression of downstream genes IFN ⁇ , IFN ⁇ 4 and CXCL10 of the cGAS-STING signal pathway, thereby activating the signal pathway.
  • the pharmacologically acceptable salt of the present invention includes inorganic acid, nitric acid, sulfuric acid, phosphoric acid, hydrobromic acid, or organic acid maleic acid, fumaric acid, tartaric acid, lactic acid, citric acid, acetic acid, methanesulfonic acid , P-toluenesulfonic acid, adipic acid, palmitic acid, tannic acid, or alkali metal lithium, sodium, potassium, or alkaline earth metal calcium, magnesium, or a salt formed by the basic amino acid lysine.
  • the present invention also discloses a pharmaceutical preparation, which contains a therapeutically effective amount of the above Rubiaceae type cyclic peptide compound or a pharmacologically acceptable salt thereof, and a pharmaceutically acceptable carrier.
  • the pharmaceutical preparation preferably contains the active ingredient Rubiaceae type cyclic peptide or its pharmacologically acceptable salt in a weight ratio of 0.1%-99.5%, and most preferably contains the active ingredient in a weight ratio of 0.5%-95%.
  • the dosage of the drug of the present invention can be changed according to the route of administration, the age, weight of the patient, the type and severity of the disease to be treated, etc.
  • the daily dose can be 0.01-10 mg/kg body weight, preferably 0.1-5 mg/kg body weight , Can be applied one or more times.
  • the pharmaceutically acceptable carrier refers to the conventional pharmaceutical carrier in the pharmaceutical field, diluent, excipient water, filler starch or sucrose; binder cellulose derivative, alginate, gelatin or polyvinylpyrrolidone; wetting agent Glycerin; disintegrant agar, calcium carbonate or sodium bicarbonate; absorption enhancer quaternary ammonium compound; surfactant cetyl alcohol; adsorption carrier kaolin or soap clay; lubricant talc, calcium stearate, magnesium stearate Or polyethylene glycol; further add other adjuvant flavors or sweeteners.
  • the pharmaceutical dosage forms of the present invention can be tablets, capsules, oral liquids, injections, freeze-dried preparations for injection, or powder injections. Because the Rubiaceae type cyclic peptides can be extracted and isolated from Rubia serrata, Rubia serrata, Rubia macrophylla, purple ginseng and other madder plants, and pharmaceutical dosage forms such as tablets, capsules, oral liquids, injections, freeze-dried preparations for injection or powder injections The preparation of is also common knowledge in the field. Therefore, various pharmaceutical dosage forms prepared from Rubiaceae type cyclic peptide compounds and corresponding carriers can also be realized by those skilled in the art.
  • the compound of the present invention can be administered to patients in need of such treatment in the form of a composition by oral, nasal inhalation, rectal or parenteral administration.
  • oral administration it can be made into conventional solid preparations such as tablets, powders, granules, capsules, etc., into liquid preparations such as water or oil suspensions or other liquid preparations such as syrups, elixirs, etc.; for parenteral use
  • liquid preparations such as water or oil suspensions or other liquid preparations such as syrups, elixirs, etc.
  • parenteral use When administered, it can be made into a solution for injection, water or oily suspension, etc.
  • Various dosage forms of the pharmaceutical composition of the present invention can be prepared according to conventional production methods in the pharmaceutical field. For example, the active ingredient is mixed with one or more carriers and then prepared into the desired dosage form.
  • the present invention detects the influence of Rubiaceae type cyclic peptides on the downstream gene expression of cGAS-STING signal pathway in HFF cells (human skin fibroblasts), MEF cells (mouse embryo fibroblasts), and Sting -/- MEF cells. And to evaluate its anti-tumor activity in vivo in colon cancer cells HT29 with normal expression of this pathway and colon cancer cells SW620 with lack of expression. It was found that the Rubiaceae type cyclic peptides can activate the human and murine cGAS-STING signaling pathways, are activators of this pathway, and can also be used to prepare drugs for the treatment and prevention of cancers related to this pathway.
  • the present invention discloses for the first time that the Rubiaceae type cyclic peptide compound is a cGAS-STING signal pathway activator, which can effectively activate the human and mouse cGAS-STING signal pathway. Therefore, it can be applied to the preparation of drugs for the treatment and prevention of tumors related to the cGAS-STING signal pathway. Because the Rubiaceae type cyclic peptide compound of the present invention is a natural compound, its dosage form and medication mode are diversified, and it has a wide range of clinical application prospects.
  • Fig. 1 shows that the Rubiaceae type cyclic peptide compound 1-27 of the present invention is used for the cGAS-STING signaling pathway downstream genes IFN ⁇ , CXCL10 and IFN ⁇ 4 mRNA of HFF cells (human skin fibroblasts) and MEF cells (mouse embryo fibroblasts) The expression of influence;
  • Figure 2 shows the effect of Rubiaceae type cyclic peptides RA-V(4) and RA-XII(9) on Ifn ⁇ mRNA in MEF and Sting -/- MEF cells;
  • Figure 3 shows the in vivo anti-tumor activity evaluation of the Rubiaceae type cyclic peptide RA-V(4) on human colon cancer cells HT29 with normal expression of cGAS-STING signaling pathway and human colon cancer cells SW620 with lack of expression.
  • the equipment, materials, and reagents used in this embodiment can be purchased through the market unless otherwise specified.
  • the preparation method of the Rubiaceae type cyclic peptide compound 1-27 described in the present invention can refer to Hu, YY, et al. Rubipodanin B, a new cytotoxic cyclopeptide from Rubia podantha.
  • Rubiaceae type cyclic peptide compounds 1-27 affect the expression of downstream genes IFN ⁇ , CXCL10 and IFN ⁇ 4 mRNA in the cGAS-STING signaling pathway in HFF cells (human skin fibroblasts) and MEF cells (mouse embryonic fibroblasts).
  • HFF cells human skin fibroblasts
  • MEF cells mouse embryonic fibroblasts
  • Cell culture Use DMEM (Invitrogen) medium containing 10% fetal bovine serum (Gibco) and 50U/mL penicillin and 50 ⁇ g/mL streptomycin to culture HFF or MEF cells at 37°C, 5% CO 2 , passage once every two or three days;
  • RNA extraction use 500 ⁇ L TRIzol to lyse the cells in (2), then add 100 ⁇ L CHCl 3 to extract the RNA in the lysate, centrifuge at 12000g at 4°C for 15min, transfer 200 ⁇ L of the supernatant to a new EP tube, add Precipitate RNA with an equal volume of isopropanol. After standing for 10 minutes, centrifuge at 12000g at 4°C for 10 minutes. Remove the supernatant. Add 1 mL of DEPC water containing 75% ethanol to wash the precipitate. Centrifuge at 7500g at 4°C for 5 min. Remove the supernatant. Dry the precipitation at room temperature for 5 minutes, and then dissolve RNA with an appropriate amount of DEPC water for subsequent experiments;
  • Real-time fluorescent quantitative PCR to detect downstream gene expression extract total cellular RNA from (2), and obtain cDNA by reverse transcription.
  • FIG. 1 The experimental results are shown in Figure 1.
  • Figures 1a, 1b, and 1c respectively show the effects of the Rubiaceae type cyclic peptide compound 1-27 of the present invention on the expression of IFN ⁇ , CXCL10 and IFN ⁇ 4 mRNA in HFF cells;
  • Figures 1d, 1e, and 1f are respectively The Rubiaceae type cyclic peptide compound 1-27 of the present invention affects the expression of Ifn ⁇ , Cxcl10 and Ifn ⁇ 4 mRNA in MEF cells.
  • MEF cells normally express cGAS-STING signaling pathway, but this pathway is absent in Sting -/- MEF cells. Therefore, by comparing the effects of the compounds on the downstream genes of the cGAS-STING pathway in these two cell lines, it can be determined whether the compound is an activator of the pathway.
  • Figure 2a shows the effect of RA-V(4) on the expression of Ifn ⁇ in MEF and Sting -/- MEF cells
  • Figure 2b shows the effect of RA-XII(9) on MEF and Sting -/- MEF cells Influence on the expression of Ifn ⁇ .
  • the experimental results show that both RA-V(4) and RA-XII(9) can effectively activate Ifn ⁇ mRNA expression in MEF cells, but their activation effects are significantly blocked in Sting -/- MEF cells, indicating that RA-V( 4) and RA-XII(9) are activators of cGAS-STING signaling pathway.
  • Rubiaceae type cyclic peptide RA-V(4) evaluates the anti-tumor activity of human colon cancer cells HT29 with normal expression of cGAS-STING signal pathway and human colon cancer cells SW620 with lack of expression in vivo.
  • Figure 3a and Figure 3b show the tumor inhibition of 2.5 mg/kg RA-V(4) on HT29 and SW620 nude mice transplanted tumor mouse models.
  • the experimental results showed that the tumor inhibition rates of 2.5 mg/kg RA-V(4) on the HT29 and SW620 nude mice transplanted tumor mouse models were 36.7% and 18.9%, respectively. It can be seen that RA-V(4) can enhance its anti-tumor activity in vivo by activating the cGAS-STING signaling pathway.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Hematology (AREA)
  • Oncology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

本发明公开了一类茜草科类型环肽化合物或其药理学上容许的盐在制备cGAS-STING信号通路激活剂的药物中的应用，以及茜草科类型环肽化合物或其药理学上容许的盐作为cGAS-STING信号通路激活剂在制备治疗和预防相关癌症药物中的应用。

Description

茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用 技术领域
本发明属于药物技术，具体地涉及一类茜草科类型环肽在制备cGAS-STING信号通路激活剂的药物中的应用。
背景技术
天然免疫是宿主防御病原微生物入侵的第一道防线，宿主细胞的模式识别受体能够识别病原微生物的保守组分病原相关分子模式，如核酸分子，感知病原微生物的入侵，通过相关信号转导级联反应，最终诱导I型干扰素以及炎症因子的表达，最终清除入侵的病原微生物。“非己”RNA识别及其识别后的信号转导机制已被详细解析，而“非己”DNA的识别信号转导机制的研究近些年才开始。
cGAS-STING信号通路是细胞内识别DNA的关键信号通路，对于干扰素及相关炎症因子的产生有着重要作用，其有助于机体识别和清除入侵的DNA病原微生物。在“非己”DNA的刺激下，cGAS迅速识别DNA，利用ATP和GTP合成cGAMP，并被内质网上的节点分子STING识别，随后迅速二聚化并从内质网转移到核外周小体上；与此同时，TBK1被招募并转移到核外周小体上激活，进而磷酸化转录因子IRF3；活化的IRF3发生二聚化并入核，激活下游干扰素及相关靶基因表达，参与抗病毒、炎症反应、免疫应答等多种生物效应。该通路与多种疾病的发生发展也有着密切关系，如肿瘤、Aicardi-Goutieres综合征和系统性红斑狼疮或狼疮样疾病。因此，在药物研发中，cGAS-STING信号通路小分子化合物调节剂的发现，包括激活剂或者抑制剂，是极有前景的研究方向。
现有技术中，能同时激活人源和鼠源cGAS-STING信号通路的小分子化合物报道很少。关于茜草科类型环肽作为cGAS-STING信号通路激活剂的制药应用，以及其作为该通路激活剂在制备治疗和预防相关癌症药物中的应用均未见报道。
发明内容
发明目的：针对上述现有技术，本申请提供了一类茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用。
技术方案：本申请公开了茜草科类型环肽化合物或其药理学上容许的盐在制备cGAS-STING信号通路激活剂的药物中的应用。
其中，所述茜草科类型环肽化合物是一类双环均环六肽类化合物，由一个D- 型α-丙氨酸、一个L-型α-丙氨酸、三个L-型N取代α-酪氨酸和一个其它L-型编码的α-氨基酸以肽链相连形成环六肽，6个氨基酸缩合成十八元环。
进一步优选的，所述茜草科类型环肽化合物为下述结构式所示的茜草科类型环肽1-27：
Figure PCTCN2021090253-appb-000001
Figure PCTCN2021090253-appb-000002
本申请所述茜草科类型环肽化合物提取自茜草属植物根及根茎。
进一步的，上述药物可用于预防和治疗cGAS-STING信号通路相关疾病，所述疾病为癌症，包括结肠癌、直肠癌、基底细胞癌、髓母细胞瘤、横纹肌肉瘤、痣样基底细胞癌综合征、小细胞肺癌、非小细胞肺癌、转移性前列腺癌、胰腺癌、软骨肉瘤、骨肉瘤、黑色素瘤、神经胶质瘤、乳腺癌、卵巢癌、食道癌、胃癌、胆管癌、肝癌、膀胱癌、血管瘤、慢性髓细胞性白血病、急性淋巴细胞性白血病、多发性骨髓瘤、霍奇金淋巴瘤、非霍奇金淋巴瘤。
所述茜草科类型环肽化合物或其药理学上容许的盐能够激活cGAS-STING信号通路下游基因IFNβ、IFNα4和CXCL10的表达，从而激活该条信号通路。
本发明所述药理学上容许的盐包括与无机酸盐酸、硝酸、硫酸、磷酸、氢溴酸，或者有机酸马来酸、富马酸、酒石酸、乳酸、柠檬酸、乙酸、甲磺酸、对苯甲磺酸、己二酸、棕榈酸、单宁酸，或者碱金属锂、钠、钾，或者碱土金属钙、镁，或者碱性氨基酸赖氨酸形成的盐。
本发明还公开了一种药物制剂，其中含有治疗有效量的上述茜草科类型环肽化合物或其药理学上容许的盐，和药学上可接受的载体。所述药物制剂中，优选含有重量比为0.1％～99.5％的活性成分茜草科类型环肽或其药理学上容许的盐，最优选含有重量比为0.5％～95％的活性成分。
本发明所述药物的施用量可根据用药途径、患者的年龄、体重、所治疗的疾病的类型和严重程度等变化，其日剂量可以是0.01～10mg/kg体重，优选0.1～5mg/kg体重，可以一次或多次施用。
所述药学上可接受的载体是指药学领域常规的药物载体，稀释剂、赋形剂水，填充剂淀粉或蔗糖；黏合剂纤维素衍生物、藻酸盐、明胶或聚乙烯吡咯烷酮；湿 润剂甘油；崩解剂琼脂、碳酸钙或碳酸氢钠；吸收促进剂季铵化合物；表面活性剂十六烷醇；吸附载体高岭土或皂黏土；润滑剂滑石粉、硬脂酸钙、硬脂酸镁或聚乙二醇；进一步加入其它辅剂香味剂或甜味剂。
本发明的药物剂型可以为片剂、胶囊、口服液、针剂、注射用冻干剂或粉针剂等。由于茜草科类型环肽可从茜草、柄花茜草、大叶茜草、紫参等茜草属植物中提取分离，而片剂、胶囊、口服液、针剂、注射用冻干剂或粉针剂等药物剂型的制备也是本领域的常规知识。因此，由茜草科类型环肽化合物与相应载体制备的各种药物剂型也能够由本领域技术人员实现。
本发明化合物可以以组合物的形式通过口服、鼻吸入、直肠或肠胃外给药的方式施用于需要这种治疗的患者。用于口服时，可将其制成常规的固体制剂如片剂、粉剂、粒剂、胶囊等，制成液体制剂如水或油悬浮剂或其他液体制剂如糖浆、酏剂等；用于肠胃外给药时，可将其制成注射用的溶液、水或油性悬浮剂等。本发明药物组合物的各种剂型可以按照药学领域的常规生产方法制备。例如使活性成分与一种或多种载体混合，然后将其制成所需的剂型。
本发明通过在HFF细胞(人皮肤成纤维细胞)、MEF细胞(小鼠胚胎成纤维细胞)、Sting -/-MEF细胞中检测茜草科类型环肽对cGAS-STING信号通路下游基因表达的影响，以及评价其对该通路正常表达的结肠癌细胞HT29和缺失表达的结肠癌细胞SW620的体内抗肿瘤活性。结果发现，茜草科类型环肽能激活人源和鼠源cGAS-STING信号通路，是该通路的激活剂，也可用于制备治疗和预防该通路相关癌症的药物。
有益效果：本发明首次揭示了茜草科类型环肽化合物是一种cGAS-STING信号通路激活剂，能够有效激活人源和鼠源cGAS-STING信号通路。由此可应用于制备治疗和预防cGAS-STING信号通路相关肿瘤的药物。由于本发明所述的茜草科类型环肽化合物为天然化合物，其剂型和用药方式多样化，具有广泛的临床应用前景。
附图说明
图1为本发明的茜草科类型环肽化合物1-27对HFF细胞(人皮肤成纤维细胞)和MEF细胞(小鼠胚胎成纤维细胞)的cGAS-STING信号通路下游基因IFNβ、CXCL10和IFNα4 mRNA的表达影响；
图2为茜草科类型环肽RA-V(4)和RA-XII(9)对MEF和Sting -/-MEF细胞中Ifnβ mRNA的影响；
图3为茜草科类型环肽RA-V(4)对cGAS-STING信号通路正常表达的人结肠癌细胞HT29和缺失表达的人结肠癌细胞SW620的体内抗肿瘤活性评价。
具体实施方式
下面结合具体实施例对本发明作出详细说明。
本实施例中使用的设备、材料、试剂除有特殊说明以外，均可以通过市场购买获取。
本发明中所述的茜草科类型环肽化合物1-27的制备方法可参照Hu,Y.Y.,et al.Rubipodanin B,a new cytotoxic cyclopeptide from Rubia podantha.Chemisty&Biodiversity,2019,16,e1800438；Chen,X.Q.,et al.Rubicordins A-C,new cyclopeptides from Rubia cordifolia with cytotoxicity and inhibiting NF-κB signaling pathway.Tetrahedron,2015,71,9673-9678；Wang,Z.,et al.Rubipodanin A,the first natural N-desmonomethyl Rubiaceae-type cyclopeptide from Rubia podantha,indicating an important role of the N 9-methyl group in the conformation and bioactivity.PLoS ONE,2015,10,e0144950；Huang,M.B.,et al.Rubischumanins A-C,three cytotoxic cyclopeptides from Rubia schumanniana.Tetrahedron,2014,70,7627-7631；Fan,J.T.,et al.Rubiyunnanins A and B,two novel cyclic hexapeptides from Rubia yunnanensis.Tetrahedron Letters,2010,51,6810-6813.Fan,J.T.,et al.Rubiyunnanins C–H,cytotoxic cyclic hexapeptides from Rubia yunnanensis inhibiting nitric oxide production and NF-κB activation.Bioorganic&Medicinal Chemistry,2010,18,8226-8234.
实施例1
茜草科类型环肽化合物1-27对HFF细胞(人皮肤成纤维细胞)和MEF细胞(小鼠胚胎成纤维细胞)的cGAS-STING信号通路下游基因IFNβ、CXCL10和IFNα4 mRNA的表达影响。实验原理、方法和结果如下：
实验原理：在固有免疫信号转导通路中，激活cGAS-STING信号通路能够诱导下游I型干扰素基因和干扰素刺激基因的表达。因此，通过检测通路下游基因IFNβ、CXCL10和IFNα4 mRNA的表达，可以评价化合物对cGAS-STING信号通路激活的影响。
实验方法：
(1)细胞培养：使用含有10％胎牛血清(Gibco)及含50U/mL盘尼西林和50μg/mL链霉素的DMEM(Invitrogen)培养基培养HFF或MEF细胞，条件为37℃、5％的CO 2，每两天或三天传代一次；
(2)用分别含10μM不同的化合物或等体积的DMSO培养基处理细胞，12小时后弃去培养液，用PBS将细胞离心收集于EP管中；
(3)RNA抽提：用500μL TRIzol将(2)中细胞裂解，然后加入100μL CHCl 3萃取裂解液中的RNA，4℃12000g离心15min，转移200μL最上层清液到新的EP管中，加入等体积的异丙醇将RNA沉淀出来，静置10min后，4℃12000g离心10min，去掉上清，加入1mL含75％乙醇的DEPC水清洗沉淀，4℃7500g离心5min，去掉上清，并在室温干燥沉淀5min，然后用适量的DEPC水溶解RNA进行后续实验；
(4)实时荧光定量PCR检测下游基因表达：以(2)中抽提得到细胞总RNA，通过逆转录得cDNA。使用Power SYBR GREEN PCR MASTER MIX(ABI)试剂进行实时荧光定量PCR，以GADPH作为内参基因。
实验结果如图1，其中图1a、1b、1c分别为本发明的茜草科类型环肽化合物1-27在HFF细胞中对IFNβ、CXCL10和IFNα4 mRNA的表达影响；图1d、1e、1f分别为本发明的茜草科类型环肽化合物1-27在MEF细胞中对Ifnβ、Cxcl10和Ifnα4 mRNA的表达影响。实验结果显示，茜草科类型环肽化合物能激活人源和鼠源细胞中cGAS-STING信号通路下游基因IFNβ，CXCL10和IFNα4的表达，其中RA-V、RA-VII和RA-III效果最好。
实施例2
茜草科类型环肽RA-V(4)和RA-XII(9)对MEF和Sting -/-MEF细胞IfnβmRNA的影响。
实验原理、方法和结果如下：
实验原理：MEF细胞正常表达cGAS-STING信号通路，而在Sting -/-MEF细胞中缺失该通路。因此，通过比较化合物对这两株细胞的cGAS-STING通路下游基因的影响，可以确定化合物是否为该通路的激活剂。
实验方法：参见实施例1。
实验结果如图2，其中图2a为RA-V(4)在MEF和Sting -/-MEF细胞中对Ifnβ 的表达影响；图2b为RA-XII(9)在MEF和Sting -/-MEF细胞中对Ifnβ的表达影响。实验结果显示，RA-V(4)和RA-XII(9)均能有效激活MEF细胞中IfnβmRNA表达，但在Sting -/-MEF细胞中其激活效果均被明显阻断，说明RA-V(4)和RA-XII(9)为cGAS-STING信号通路的激活剂。
实施例3
茜草科类型环肽RA-V(4)对cGAS-STING信号通路正常表达的人结肠癌细胞HT29和缺失表达的人结肠癌细胞SW620的体内抗肿瘤活性评价。
实验方法和结果如下：
将人结肠癌细胞HT29和SW620用生理盐水稀释成2×10 7个/mL，取100μL该细胞悬浮液接种于BABL/c裸鼠左侧腋窝皮下，生长7天，形成荷瘤小鼠模型。取接种生长良好的荷瘤小鼠，随机分组，取RA-V(4)纳米胶束注射剂按2.5mg/kg通过尾静脉给药，隔天给药一次并测量体积，给药12天后处死所用动物，计算抑瘤率，统计处理。抑瘤率(％)＝(对照组平均体积-实验组平均体积)/对照组平均体积×100％。
实验结果见图3，其中图3a和图3b分别为2.5mg/kg RA-V(4)在HT29和SW620裸鼠移植瘤小鼠模型上抑瘤情况。实验结果显示，2.5mg/kg RA-V(4)在HT29和SW620裸鼠移植瘤小鼠模型上的抑瘤率分别为36.7％和18.9％。可见，RA-V(4)可通过激活cGAS-STING信号通路增强其体内抗肿瘤活性。

Claims (9)

  1. 茜草科类型环肽化合物或其药理学上容许的盐在制备cGAS-STING信号通路激活剂的药物中的应用。
  2. 根据权利要求1所述的应用，其特征在于，所述茜草科类型环肽化合物是一类双环均环六肽类化合物，由一个D-型α-丙氨酸、一个L-型α-丙氨酸、三个L-型N取代α-酪氨酸和一个其它L-型编码的α-氨基酸以肽链相连形成环六肽，6个氨基酸缩合成十八元环。
  3. 根据权利要求1所述的应用，其特征在于，所述茜草科类型环肽化合物为下述结构式所示的茜草科类型环肽1-27：
    Figure PCTCN2021090253-appb-100001
    Figure PCTCN2021090253-appb-100002
  4. 根据权利要求1-3中任一所述的应用，其特征在于，所述茜草科类型环肽化合物提取自茜草属植物根及根茎。
  5. 根据权利要求1-3中任一所述的应用，其特征在于，所述药物可用于预防和治疗cGAS-STING信号通路相关疾病，所述疾病为癌症，包括结肠癌、直肠癌、基底细胞癌、髓母细胞瘤、横纹肌肉瘤、痣样基底细胞癌综合征、小细胞肺癌、非小细胞肺癌、转移性前列腺癌、胰腺癌、软骨肉瘤、骨肉瘤、黑色素瘤、神经胶质瘤、乳腺癌、卵巢癌、食道癌、胃癌、胆管癌、肝癌、膀胱癌、血管瘤、慢性髓细胞性白血病、急性淋巴细胞性白血病、多发性骨髓瘤、霍奇金淋巴瘤、非霍奇金淋巴瘤。
  6. 根据权利要求5所述的应用，其特征在于，所述茜草科类型环肽化合物或其药理学上容许的盐能够激活cGAS-STING信号通路下游基因IFNβ、IFNα4和CXCL10的表达。
  7. 根据权利要求1所述的应用，其特征在于，所述药理学上容许的盐包括与无机酸盐酸、硝酸、硫酸、磷酸、氢溴酸，或者有机酸马来酸、富马酸、酒石酸、乳酸、柠檬酸、乙酸、甲磺酸、对苯甲磺酸、己二酸、棕榈酸、单宁酸，或者碱金属锂、钠、钾，或者碱土金属钙、镁，或者碱性氨基酸赖氨酸形成的盐。
  8. 一种药物制剂，其中含有治疗有效量的权利要求1所述的茜草科类型环肽化合物或其药理学上容许的盐，和药学上可接受的载体。
  9. 根据权利要求8所述的药物制剂，其特征在于，所述茜草科类型环肽化合物或其药理学上容许的盐的含量为0.1％～99.5％。
PCT/CN2021/090253 2020-04-28 2021-04-27 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用 WO2021218965A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202010347027.3 2020-04-28
CN202010347027.3A CN111420025B (zh) 2020-04-28 2020-04-28 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用

Publications (1)

Publication Number Publication Date
WO2021218965A1 true WO2021218965A1 (zh) 2021-11-04

Family

ID=71552007

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2021/090253 WO2021218965A1 (zh) 2020-04-28 2021-04-27 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用

Country Status (2)

Country Link
CN (1) CN111420025B (zh)
WO (1) WO2021218965A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114478700A (zh) * 2022-02-22 2022-05-13 南京医科大学 鸡冠花子中荨麻科类型环肽的制备方法及其在抗肿瘤药物中的应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111420025B (zh) * 2020-04-28 2021-06-11 中国药科大学 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103877562A (zh) * 2014-04-03 2014-06-25 中国科学院昆明植物研究所 茜草科类型环肽用作为Hedgehog信号通路抑制剂和其制备方法与应用
CN103880929A (zh) * 2014-04-03 2014-06-25 中国科学院昆明植物研究所 茜草科类型环肽用作为肿瘤转移抑制剂和其制备方法与应用
CN104193808A (zh) * 2014-09-03 2014-12-10 中国科学院昆明植物研究所 茜草科类型环肽制备方法及其用作为Hippo-YAP信号通路抑制剂
CN104829696A (zh) * 2015-05-12 2015-08-12 中国药科大学 开环茜草科类型环肽,以其为活性成分的药物组合物,其制备方法和应用
CN107496901A (zh) * 2017-10-12 2017-12-22 中国药科大学 细胞自噬抑制剂及其制备方法与应用
CN107970432A (zh) * 2017-12-05 2018-05-01 中国药科大学 以植物环肽为有效成分的肿瘤细胞异常脂代谢抑制剂及其应用
CN111420025A (zh) * 2020-04-28 2020-07-17 中国药科大学 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101921319B (zh) * 2010-07-21 2013-01-09 中国科学院昆明植物研究所 茜草科类型环肽,以其为活性成分的药物组合物,其制备方法和应用
CN104262465B (zh) * 2014-09-03 2017-01-11 中国科学院昆明植物研究所 茜草科类型环肽用作为tak1抑制剂和其制备方法
JP6898848B2 (ja) * 2014-09-19 2021-07-07 メモリアル スローン ケタリング キャンサー センター 脳転移を治療するための方法
EP4339287A3 (en) * 2015-07-31 2024-05-22 Regents Of The University Of Minnesota Modified cells and methods of therapy
WO2018081476A2 (en) * 2016-10-27 2018-05-03 Intima Bioscience, Inc. Viral methods of t cell therapy
CN107335049B (zh) * 2017-08-18 2019-10-18 中国药科大学 菊科类型环肽化合物作为cGAS-STING信号通路抑制剂的应用

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103877562A (zh) * 2014-04-03 2014-06-25 中国科学院昆明植物研究所 茜草科类型环肽用作为Hedgehog信号通路抑制剂和其制备方法与应用
CN103880929A (zh) * 2014-04-03 2014-06-25 中国科学院昆明植物研究所 茜草科类型环肽用作为肿瘤转移抑制剂和其制备方法与应用
CN104193808A (zh) * 2014-09-03 2014-12-10 中国科学院昆明植物研究所 茜草科类型环肽制备方法及其用作为Hippo-YAP信号通路抑制剂
CN104829696A (zh) * 2015-05-12 2015-08-12 中国药科大学 开环茜草科类型环肽,以其为活性成分的药物组合物,其制备方法和应用
CN107496901A (zh) * 2017-10-12 2017-12-22 中国药科大学 细胞自噬抑制剂及其制备方法与应用
CN107970432A (zh) * 2017-12-05 2018-05-01 中国药科大学 以植物环肽为有效成分的肿瘤细胞异常脂代谢抑制剂及其应用
CN111420025A (zh) * 2020-04-28 2020-07-17 中国药科大学 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEN JUANJUAN, ZHANG PINGAN: "Advances in cGAS-STING Signaling Pathway in Cancer", AIZHENG JINZHAN = ONCOLOGY PROGRESS, ZHONGGUO XIEHE YIKE DAXUE CHUBANSHE, CN, vol. 17, no. 4, 25 February 2019 (2019-02-25), CN , XP055861052, ISSN: 1672-1535, DOI: 10.11877/j.issn.1672-1535.2019.17.04.03 *
FAN, J.T. ; CHEN, Y.S. ; XU, W.Y. ; DU, L. ; ZENG, G.Z. ; ZHANG, Y.M. ; SU, J. ; LI, Y. ; TAN, N.H.: "Rubiyunnanins A and B, two novel cyclic hexapeptides from Rubia yunnanensis", TETRAHEDRON LETTERS, ELSEVIER, AMSTERDAM , NL, vol. 51, no. 52, 29 December 2010 (2010-12-29), Amsterdam , NL , pages 6810 - 6813, XP027516784, ISSN: 0040-4039, DOI: 10.1016/j.tetlet.2010.07.066 *
FAN, J.T. ; SU, J. ; PENG, Y.M. ; LI, Y. ; LI, J. ; ZHOU, Y.B. ; ZENG, G.Z. ; YAN, H. ; TAN, N.H.: "Rubiyunnanins C-H, cytotoxic cyclic hexapeptides from Rubia yunnanensis inhibiting nitric oxide production and NF-@kB activation", BIOORGANIC, ELSEVIER, AMSTERDAM, NL, vol. 18, no. 23, 1 December 2010 (2010-12-01), AMSTERDAM, NL, pages 8226 - 8234, XP027546679, ISSN: 0968-0896 *
ZHE WANG, SIMENG ZHAO, LIHUA SONG, YUZHI PU, QIANG WANG, GUANGZHI ZENG, XING LIU, MING BAI, SHAO LI, FABAO GAO, LIJUAN CHEN, CHEN : "Natural cyclopeptide RA-V inhibits the NF-κB signaling pathway by targeting TAK1", CELL DEATH & DISEASE, vol. 9, no. 7, 1 July 2018 (2018-07-01), XP055718053, DOI: 10.1038/s41419-018-0743-2 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114478700A (zh) * 2022-02-22 2022-05-13 南京医科大学 鸡冠花子中荨麻科类型环肽的制备方法及其在抗肿瘤药物中的应用
CN114478700B (zh) * 2022-02-22 2023-09-08 南京医科大学 鸡冠花子中荨麻科类型环肽的制备方法及其在抗肿瘤药物中的应用

Also Published As

Publication number Publication date
CN111420025B (zh) 2021-06-11
CN111420025A (zh) 2020-07-17

Similar Documents

Publication Publication Date Title
WO2021213288A1 (zh) 核苷类似物或含有核苷类似物的组合制剂在抗病毒中的应用
JP5694345B2 (ja) Ｔｏｌｌ様受容体の調節因子
WO2021218965A1 (zh) 茜草科类型环肽化合物在制备cGAS-STING信号通路激活剂的药物中的应用
CN111557939B (zh) 法匹拉韦在制备用于治疗冠状病毒感染的药物中的应用
WO2017162108A1 (zh) 一种柱芳烃类复合物、其制备方法、药物组合物和用途
WO2019033710A1 (zh) 菊科类型环肽化合物作为cGAS-STING信号通路抑制剂的应用
JP2009517403A (ja) Ｐａｒｐ−１阻害剤の使用
Hu et al. Japonicone A antagonizes the activity of TNF-α by directly targeting this cytokine and selectively disrupting its interaction with TNF receptor-1
CN105246887A (zh) 香豆素衍生物以及用于治疗过度增生性疾病的方法
US20170360743A1 (en) Biflavone compound and uses thereof for treating cancers and preparing drugs
WO2019227764A1 (zh) 一种mln4924在制备治疗糖尿病和/或肥胖药物的新用途
RU2657604C2 (ru) Противоопухолевый агент, включающий гидрат гидрохлорида иринотекана
TW202024117A (zh) 抗紅血球生成素受體胜肽
CN107324999B (zh) 萘醌二聚体及其制备方法与应用
WO2018058863A1 (zh) 聚醚类化合物用途
WO2017045595A1 (zh) 环二核苷酸cGAMP联合贝伐珠单抗在抗肿瘤中的应用
CN115190800A (zh) 一种brd4抑制剂的用途
CN102319260A (zh) 顺铂联合伊曲康唑异构体在制备治疗肺癌药物中的应用
WO2019101040A1 (zh) 含有双环铂的组合产品、其制备方法及其用途
CN114805141A (zh) 4-胍基苯甲酸芳基酯类化合物及其在抗SARS-CoV-2病毒中的用途
CN107773762B (zh) 基于pd-l1抗体偶联化疗药物的adc及其制备方法和应用
CN106905275B (zh) 一种4-芳基吡喃衍生物及其制备和应用
CN111821303A (zh) 沃替西汀及其盐在制备抗肿瘤药物中的应用
CN110403924A (zh) 一种治疗皮肤黑色素瘤的药物组合物及其制备方法
WO2007062528A1 (en) Polycyclic phenolic compounds and use in treating viral infections

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 21796628

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 21796628

Country of ref document: EP

Kind code of ref document: A1