WO2024027673A1 - 红色诺卡氏菌细胞壁骨架在治疗慢性宫颈炎中的应用 - Google Patents

红色诺卡氏菌细胞壁骨架在治疗慢性宫颈炎中的应用 Download PDF

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WO2024027673A1
WO2024027673A1 PCT/CN2023/110460 CN2023110460W WO2024027673A1 WO 2024027673 A1 WO2024027673 A1 WO 2024027673A1 CN 2023110460 W CN2023110460 W CN 2023110460W WO 2024027673 A1 WO2024027673 A1 WO 2024027673A1
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nocardia
cell wall
rubrum
wall skeleton
cervical
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PCT/CN2023/110460
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English (en)
French (fr)
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盖波
张轶
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辽宁天安生物制药股份有限公司
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

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  • the present disclosure relates to the fields of medicine and biopharmaceuticals. Specifically, the present disclosure relates to the use of Nocardia rubrum cell wall scaffolds in the treatment of chronic cervicitis.
  • Chronic cervicitis is the most common disease among women of childbearing age and is often caused by Streptococcus, Enterococcus, Escherichia coli and Staphylococcus aureus. Special pathogenic microorganisms include Chlamydia trachomatis and Neisseria gonorrhoeae. In addition, childbirth and mechanical injury are also predisposing factors for chronic cervicitis.
  • Nocardia rubra is a type of Nocardia.
  • the Nocardia rubrum cell wall skeleton can be obtained by fermentation, cell disruption and protease degradation of the Nocardia rubrum cells.
  • Nocardia rubrum the cell wall skeleton of Nocardia rubrum can be commercially available, specifically a product produced by Liaoning Greenstone Biopharmaceutical Co., Ltd.
  • Nocardia rubrum cell wall skeleton has been used to treat cervical precancerous lesions (CN101073583A), against human papillomavirus (CN1935262A), skin lesions or skin ulcers (CN101209267A), fungal infections, herpes simplex, and herpes zoster (CN1879661A).
  • the present disclosure provides a Nocardia rubrum cell wall scaffold.
  • Nocardia rubra refers to Nocardia rubra species of the genus Nocardia.
  • Identification of Nocardia rubrum Based on known or future microbial identification technologies, technicians can conduct taxonomic identification of a strain of bacteria. For example, available identification technologies include morphology, physiological and biochemical characteristics, 16S rRNA, etc. Technicians understand that with the development of science and technology, identification technology involves different means. In earlier periods, morphological and biochemical identification methods were mainly used, but the reliability of this method is not high. The advent of sequencing technology has given technicians a more reliable way to identify strains. For example, when the DNA sequence of 16S rRNA is identified as having a similarity of 97% or more (inclusive), the two bacteria are determined to belong to the same species. At once For Nocardia rubrum, known strains deposited in international (or national) culture collection units are used as model strains and compared with them.
  • Nocardia rubrum cell wall can be understood as either a complete cell wall or an incomplete cell wall (for example, broken or partially degraded).
  • the component exhibiting the desired activity is derived from the cell wall of Nocardia rubrum (eg, the cell wall itself or a component thereof). Therefore, various forms such as intact cell walls, broken cell walls, incomplete cell wall degradation products, cell wall components, cell wall extracts, etc. are allowed to be used in clinical applications, all of which are included in the scope of this disclosure.
  • the cell wall skeleton of the present disclosure cannot be understood to only represent the cross-linked network entities in the cell wall. Skilled persons should understand that this term does not exclude other cell wall components that are adsorbed, combined, and carried on the cross-linked network entities.
  • the cell wall skeleton of the present disclosure is a product obtained by disrupting bacteria and removing impurities (proteins, nucleic acids, cell membranes, and lipids).
  • the cell wall skeleton is the Nocardia rubrum cell wall skeleton corresponding to National Drug Approval Number S20030009 or its equivalent standard.
  • S20030009 is the initial administrative license number issued by the drug regulatory department. This number changes with the update of certificates, adjustments to laws and numbering rules. However, no matter how the number changes, the product standards (product parameters and/or quality requirements) it represents remain unchanged. Therefore, S20030009 in this disclosure should be understood to also include: S20030009 itself, its equivalent number, the cell wall skeleton corresponding to the updated number, and also includes the cell wall skeleton with the same product parameters and/or quality requirements as S20030009.
  • Cell wall scaffold with the same product parameters and/or quality requirements or “cell wall scaffold corresponding to equivalent standards” means that under the same testing method, the cell wall scaffold to be tested and the cell wall scaffold shown in S20030009 have no statistically significant difference in activity Ingredients (muramic acids, sugars, lipids).
  • the Nocardia rubrum cell wall skeleton is obtained by the following method, which method includes or consists of the following steps:
  • Steps 3.1), 3.2), 3.3), and 3.4) can be interchanged or parallel, and steps 4) and 5) can be interchanged.
  • Nocardia rubrum disruption the purpose is to remove intracellular material. Therefore, technologies such as ultrasonic crushing, high-pressure homogenizer crushing, and lysozyme can be used.
  • any known or future method suitable for disrupting Gram-positive bacteria is applicable to the technical solution of the present disclosure.
  • organic solvents are used to remove lipids from the fragmentation product.
  • nucleases are used to remove DNA and RNA from fragmentation products.
  • hydrolytic enzymes are utilized to degrade proteins in the fragmentation product.
  • surfactants are used to remove cell membranes from the disrupted product.
  • the average particle size of the fragmentation is from 10 nm to 1000 nm; mention may be made of 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190nm ⁇ 10nm, and the range between any two of the above values. Testing methods for particle size are well known in the art.
  • the average particle size of the fragmentation is from 10 nm to 800 nm.
  • the average particle size of the fragmentation is from 10 nm to 500 nm.
  • said dispensing refers to dispensing into bottles or ampoules. Just before use, add solvent (such as sterile water) to the bottle or ampoule.
  • solvent such as sterile water
  • the vial refers to a vial (made of borosilicate glass or soda-lime glass).
  • the present disclosure provides the use of the aforementioned Nocardia rubrum cell wall skeleton in the preparation of a medicament, wherein the medicament is used to prevent or treat chronic cervicitis associated with high-risk HPV infection in a subject.
  • the high-risk HPV is selected from any one of the following or a combination thereof: types 16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, and 68.
  • the chronic cervicitis is selected from any one of the following or a combination thereof: cervical columnar epithelial ectopia, cervical polyps, cervical mucositis, cervical gland cyst, cervical hypertrophy (classification can be found in, for example, "Obstetrics and Gynecology" published by People's Health Publishing House , seventh edition).
  • cervical columnar epithelial heterotopia is the most common local feature of the inflammatory process of chronic cervicitis.
  • the red lesions on the surface of the cervix are the result of the shedding of squamous epithelium, replacement by columnar epithelium, and exposure of subepithelial blood vessels.
  • cervical polyps are one of the manifestations of chronic cervicitis. Due to long-term stimulation of chronic inflammation, local mucosa of the cervical canal proliferates. Because the uterus has a tendency to expel foreign bodies, the proliferated mucosa can gradually protrude from the base to the external os of the cervix to form polyps.
  • cervical mucositis is one of the common manifestations of chronic cervicitis.
  • the lesions are located in the cervical mucosa and submucosal tissue.
  • cervical adenocyst is one of the common manifestations of chronic cervicitis.
  • the new squamous epithelium covers the opening of the cervical gland duct or extends into the gland duct, blocking the gland duct opening; the connective tissue hyperplasia or scar formation around the gland duct compresses the gland duct and narrows the gland duct. Even blocked, the drainage of glandular secretions is blocked, and the cyst formed by retention is called cervical Nassler cyst. Cervical adenocysts are inflammation, not tumors.
  • cervical hypertrophy is a form of chronic cervicitis. Inflammatory changes caused by pathogens infecting the cervical mucosa.
  • the hypertrophied cervical surface may also experience squamous epithelial shedding and columnar epithelial hyperplasia due to injury or inflammatory stimulation.
  • the drug is prepared in a dosage form selected from any one of the following: injections, ointments, creams, emulsions, suspensions, pastes, gels, lotions, tablets, aerosols, Sprays, liniments, powders, dressings, bandages, films, patches, suppositories.
  • the present disclosure provides a pharmaceutical composition or medicament for preventing or treating chronic cervicitis associated with high-risk HPV infection in a subject, which includes: a pharmaceutically acceptable carrier and a medicament of the present disclosure.
  • compositions or medicaments of the present disclosure may be prepared in unit dose (or unit preparation) form.
  • a pharmaceutical composition or drug may be prepared in a liquid form (liquid formulation).
  • the pharmaceutical composition or drug can be prepared as a solid (dry powder formulation or lyophilized powder formulation).
  • liquid preparations and dry powder preparations can be converted into each other, and the only difference lies in the water content. Most or all of the water in the liquid preparation is removed to obtain a dry powder preparation (or freeze-dried powder preparation). The dry powder preparation (or freeze-dried powder preparation) is dissolved (or reconstituted) to obtain a liquid preparation.
  • the pharmaceutically acceptable carrier is selected from, but is not limited to: fillers, stabilizers (such as trehalose, glycine), flavoring agents (such as xylitol), disintegrants (such as carboxylic acid) Sodium methylcellulose), binder (e.g. gelatin), lubricant (e.g. magnesium stearate).
  • the stabilizer is selected from one or a combination of the following: glycine, lysine, arginine, hydroxyethyl starch, hydroxymethyl starch, trehalose, dextran.
  • the flavoring agent is selected from one or a combination of sucrose, monosaccharide, sodium saccharin, aspartame, sorbitol, xylitol, mannitol.
  • the binder is selected from one or a combination of sodium carboxymethylcellulose, hypromellose, and gelatin.
  • the lubricant is selected from one or a combination of talc, magnesium stearate, and micronized silica gel.
  • pharmaceutically acceptable carriers suitable for use in the present disclosure may also be mentioned, such as but not limited to: dextran, lactose, microcrystalline cellulose, trehalose, glycine, xylitol, carboxymethyl Sodium cellulose, erythritol, gelatin, magnesium stearate, propellant, humectant, solvent, solubilizer, emulsifier, antioxidant, pH adjuster, preservative.
  • non-limiting examples also include: white petrolatum, carbomer, hypromellose, methylcellulose, sodium carboxymethylcellulose, chitosan, sucralfate chitosan, polyvinylpyrrolidone, Polyvinyl alcohol, sodium hyaluronate, dimethyl ether, tetrafluoroethane, hydrofluoroalkane, glycerin, propylene glycol, deionized water, water for injection, distilled water, ethanol, cetyl alcohol, stearyl alcohol, para-aminobenzoic acid, ethyl alcohol Amide, isopropyl alcohol, Tween, polyoxyethyl hydrogenated castor oil, stearic acid, glyceryl monostearate, triglyceryl monostearate, fatty acid sucrose ester, sucrose ester, acetic acid isobutyric acid sucrose sugar Ester, sorbitan tristearate, isopropyl myristate
  • the present disclosure provides a method for preventing or treating chronic cervicitis associated with high-risk HPV infection in a subject, comprising the steps of: administering a therapeutically effective amount of Nocardia rubrum of the present disclosure to the subject Cell wall skeleton or pharmaceutical composition.
  • administer when applied to an animal, human, cell, tissue, organ or biological sample means that the drug or medical device comes into contact with the animal, human, cell, tissue, organ or biological sample.
  • Treatment means administering to a subject an internal or external drug (therapeutic agent, active ingredient or composition) (e.g., exosomes or pharmaceutical compositions of the present disclosure) or medical device, and in the subject being treated ( Alleviating (mitigating, delaying, ameliorating, curing) one or more disease symptoms to a clinically measurable extent (or group), wherein said subject has suffered from, is suspected of suffering from, or is susceptible to a disease one or more diseases or their symptoms.
  • an internal or external drug e.g., active ingredient or composition
  • therapeutic agent e.g., exosomes or pharmaceutical compositions of the present disclosure
  • the amount of a drug (therapeutic agent, active ingredient or composition) that is effective in alleviating the symptoms of any disease is called a therapeutically effective amount.
  • a drug therapeutic agent, active ingredient or composition
  • the subject is an animal other than a human, such as a farm animal, a pet, a working animal, an ornamental animal, a production animal, a laboratory animal (such as a rat, mouse, guinea pig, rabbit, dog, primates).
  • a farm animal such as a farm animal, a pet, a working animal, an ornamental animal, a production animal, a laboratory animal (such as a rat, mouse, guinea pig, rabbit, dog, primates).
  • the subject is human. In some specific embodiments, the subject is suspected of having, diagnosed with, has suffered from, or is susceptible to the target disease or symptoms thereof.
  • the administration is 1-3 times a day, or once a day, or once every other day. Depending on the area and degree of the patient's lesions, different doses are used.
  • the administration cycle lasts from 2 days to 6 months, e.g., 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks or longer , and the range between any two aforementioned values.
  • the same pharmaceutical active ingredient or different pharmaceutical active ingredients may be administered at once, or may be divided into many smaller unit doses and administered at regular intervals. It is understood that the exact dosage, duration, and intervals of treatment are a function of the disease being treated and can be determined using extrapolation from animal or clinical trial data.
  • the administration may comprise a single administration, or two or more administrations spaced at appropriate intervals.
  • two consecutive administrations can be separated by 30 minutes, 40 minutes, 50 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 12 hours hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, one and a half days, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months or 12 months.
  • Optional means that the events described subsequently may occur, but do not have to occur; it depends on the circumstances.
  • dispensed means that the product is allowed to be dispensed, but is not required to be dispensed.
  • Example 1 Commercially available cell wall scaffold of Nocardia rubrum
  • the cell wall skeleton of Nocardia rubrum was purchased from Liaoning Greenstone Biopharmaceutical Co., Ltd., with the national drug approval number S20030009 (the solid content of the cell wall skeleton in each tube should be no less than 60 ⁇ g, The muramic acid content is not less than 1.0 ⁇ g, and the sugar content is not less than 4.0 ⁇ g; the reconstitution volume is 2.0 ml).
  • Lipid removal Add organic reagents (such as one or a combination of acetone, ether, ethanol) to the precipitate, and remove lipids according to conventional operations in this field.
  • organic reagents such as one or a combination of acetone, ether, ethanol
  • Example 1 Coat the product obtained in Example 1 or 2 on a dressing (such as sterile gauze) to prepare an external medicine.
  • a dressing such as sterile gauze
  • lotions mostly use water and alcohol as the dispersion medium; they are made from active ingredients, electrolytes, isotonic regulators, etc. in the dispersion medium.
  • the cell wall scaffold of Example 1 was administered to subjects clinically diagnosed with chronic cervicitis.
  • the treatment results are shown in Table 1.

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Abstract

提供了一种红色诺卡氏菌细胞壁骨架在制备药物中的用途，其中所述药物用于在受试者中预防或治疗伴有高危型HPV感染的慢性宫颈炎。

Description

红色诺卡氏菌细胞壁骨架在治疗慢性宫颈炎中的应用 技术领域
本公开涉及医学、生物制药领域。具体而言，本公开涉及红色诺卡氏菌细胞壁骨架在治疗慢性宫颈炎中的应用。
背景技术
慢性宫颈炎为育龄期妇女最常见的疾病，常由链球菌、肠球菌、大肠杆菌和葡萄球菌引起，特殊的病原微生物包括沙眼衣原体、淋球菌。此外，分娩、机械损伤也是慢性子宫颈炎的诱发因素。
红色诺卡氏菌(Nocardia rubra)是诺卡氏菌中的一种。红色诺卡氏菌的菌体经发酵、细胞破碎、蛋白酶降解后可制得红色诺卡氏菌细胞壁骨架。
现有技术中，红色诺卡氏菌细胞壁骨架可以是市售获得，具体而言由辽宁格瑞仕特生物制药有限公司生产的商品。红色诺卡氏菌细胞壁骨架已用于治疗宫颈癌前病变(CN101073583A)、抗人乳头瘤病毒(CN1935262A)、皮肤损伤或皮肤溃疡(CN101209267A)、真菌感染、单纯疱疹、带状疱疹(CN1879661A)。
发明内容
第一方面，本公开提供了一种红色诺卡氏菌细胞壁骨架。
红色诺卡氏菌是指Nocardia属的红色诺卡氏菌种(Nocardia rubra)。
红色诺卡氏菌的鉴定：根据已知的或未来的微生物鉴定技术，技术人员可以对一株细菌进行分类学鉴定，例如可用的鉴定技术包括形态学、生理生化特征、16S rRNA等。技术人员理解，随着科技的发展，鉴定技术涉及不同的手段，在较早的时期主要采用形态学和生化鉴定方式，但是这种方法的可靠程度不高。测序技术出现后，技术人员可以利用更为可信的方式鉴定菌株。例如，当16S rRNA的DNA序列被鉴定为具有97％(含)以上相似性时，判定两个菌属于相同的种。就 红色诺卡氏菌而言，将保藏在国际(或国家级)菌种保藏单位中的已知菌株作为模式菌株，并与其进行比对。
本公开中，“红色诺卡氏菌细胞壁”既可以理解为完整的细胞壁，也可以理解为不完整的细胞壁(例如，破碎的、或部分降解的)。在本公开的教导下，技术人员将理解，显示出所需活性的成分来自红色诺卡氏菌的细胞壁(例如，是细胞壁本身或其组成成分)。因此，在临床应用中允许采用完整的细胞壁、经破碎的细胞壁、细胞壁的不完全降解产物、细胞壁的组成成分、细胞壁的提取物等各种形式，这些都包含在本公开范畴之内。
本公开的细胞壁骨架不能理解为仅仅表示细胞壁当中的交联网状实体，技术人员应理解该术语不排除交联网状实体上所吸附、结合、携带的其他细胞壁成分。
在具体的示例中，本公开的细胞壁骨架是细菌经破碎、除杂(蛋白、核酸、细胞膜、脂质)后所得的产物。
在具体的实施方案中，细胞壁骨架是国药准字S20030009或其等同标准对应的红色诺卡氏菌细胞壁骨架。
技术人员应当理解，S20030009是药监管理部门发放的行政许可初始编号，此编号随着证件的更新、法律、编号规则的调整而变化。然而，无论编号如何变化，其所代表的产品标准(产品参数和/或质量要求)是不变的。因此，本公开中S20030009应理解为还包括：S20030009本身、其等同编号、更新编号对应的细胞壁骨架，也包括与S20030009具有相同产品参数和/或质量要求的细胞壁骨架。
“具有相同产品参数和/或质量要求的细胞壁骨架”或“等同标准对应的细胞壁骨架”是指在相同的测试方法下，待测细胞壁骨架和S20030009所示细胞壁骨架具有无统计学显著差异的活性成分(胞壁酸、糖、脂质)。
在另一些实施方案中，所述红色诺卡氏菌细胞壁骨架通过以下方法获得，所述方法包括以下步骤或由以下步骤组成：
1)提供红色诺卡氏菌；
2)破碎所述红色诺卡氏菌，得到破碎产物；
3.1)对所述破碎产物去除脂质；
3.2)对所述破碎产物去除核酸；
3.3)对所述破碎产物去除蛋白质；
3.4)对所述破碎产物去除细胞膜；
3.5)得到红色诺卡氏菌细胞壁骨架；
4)任选地，分装；
5)任选地，对所述红色诺卡氏菌细胞壁骨架进行冷冻干燥；
步骤3.1)、3.2)、3.3)、3.4)能够互换或并行，步骤4)和步骤5)能够互换。
对于红色诺卡氏菌的破碎，其目的在于去除细胞内的物质。因此可以采用超声破碎、高压均质机破碎、溶菌酶等技术。技术人员理解，任何适用于破碎革兰氏阳性菌的已知或未来方法，均适用于本公开的技术方案。
技术人员有能力根据活性成分(细胞壁及其组成成分)的后续应用(例如外敷)，来调整培养、破碎、分离、收集、除杂质、分装的具体参数和设备，以免制备步骤中引入影响后续应用的因素。
在一些实施方案中，利用有机溶剂去除破碎产物中的脂质。在一些实施方案中，利用核酸酶去除破碎产物中的DNA和RNA。在一些实施方案中，利用水解酶降解破碎产物中的蛋白质。在一些实施方案中，利用表面活性剂去除破碎产物中的细胞膜。
在一些实施方案中，破碎的平均粒度为10nm至1000nm；可以提及10、20、30、40、50、60、70、80、90、100、110、120、130、140、150、160、170、180、190nm±10nm，以及上述任意两个数值之间的范围。粒度的测试方法是现有技术公知的。
在一些具体的实施方案中，破碎的平均粒度为10nm至800nm。
在另一些具体的实施方案中，破碎的平均粒度为10nm至500nm。
在具体的实施方案中，所述分装是指分装至瓶或安瓿中。临用前，向瓶或安瓿中添加溶剂(如无菌水)。作为一个示例，瓶是指西林瓶(vial，由硼硅玻璃或钠钙玻璃制成)。
第二方面，本公开提供了前述红色诺卡氏菌细胞壁骨架在制备药物中的用途，其中所述药物用于在受试者中预防或治疗伴有高危型HPV感染的慢性宫颈炎。
所述高危型HPV选自以下的任一项或其组合：16、18、31、33、35、39、45、51、52、53、56、58、59、66、68型。
所述慢性宫颈炎选自以下的任一项或其组合：宫颈柱状上皮异位、宫颈息肉、宫颈粘膜炎、宫颈腺囊肿、宫颈肥大(分类可见于例如《妇产科学》人民卫生出版社出版，第七版)。
在一些实施方案中，宫颈柱状上皮异位是慢性宫颈炎的炎性病变过程中最多见的局部特征。宫颈表面呈红色病损，是鳞状上皮脱落，为柱状上皮所代替，上皮下血管显露的结果。临床常根据病灶面积，将其分成轻I°(病灶面积小于子宫颈总面积1/3)、中II°(1/3-1/2)、重度III°(超过1/2)。
在一些实施方案中，宫颈息肉是慢性宫颈炎表现之一。由于慢性炎症长期刺激，使宫颈管局部黏膜增生，因子宫有排除异物倾向，可使增生的黏膜逐渐自基底部向宫颈外口突出而形成息肉。
在一些实施方案中，宫颈粘膜炎是慢性宫颈炎常见的表现之一。病变位于宫颈管粘膜及粘膜下组织。
在一些实施方案中，宫颈腺囊肿是慢性宫颈炎常见的表现之一。宫颈柱状上皮异位恢复过程中，新生的鳞状上皮覆盖宫颈腺管口或伸入腺管，将腺管口阻塞；腺管周围的结缔组织增生或瘢痕形成压迫腺管，使腺管变窄甚至阻塞，腺体分泌物引流受阻，滞留形成的囊肿叫宫颈纳氏囊肿。宫颈腺囊肿是炎症，而非肿瘤。
在一些实施方案中，宫颈肥大是慢性宫颈炎的一种。病原体感染宫颈粘膜引起的炎性改变。肥大的宫颈表面由于损伤或炎症刺激也会出现鳞状上皮脱落、柱状上皮增生。
在一些实施方案中，所述药物制备成选自以下任一项的剂型：注射剂、膏剂、霜剂、乳液、混悬剂、糊剂、凝胶剂、洗剂、片剂、气雾剂、喷雾剂、搽剂、粉剂、敷料、绷带、膜、贴片、栓剂。
第三方面，本公开提供了一种用于在受试者中预防或治疗伴有高危型HPV感染的慢性宫颈炎的药物组合物或药物，其包含：药学上可接受的载体和本公开的红色诺卡氏菌细胞壁骨架。
本公开的药物组合物或药物可以制备成单位剂量(或单元制剂)的形式。
在一些实施方案中，药物组合物或药物可以制备成液态的(液体制剂)。
在另一些实施方案中，药物组合物或药物可以制备成固体的(干粉制剂或冻干粉制剂)。
技术人员理解，液体制剂和干粉制剂(或冻干粉制剂)，二者可以相互转化，差别仅在于含水量。除去液体制剂中的绝大部分或全部水，得到干粉制剂(或冻干粉制剂)。干粉制剂(或冻干粉制剂)溶解(或复溶)后得到液体制剂。
在一些实施方案中，所述药学上可接受的载体选自，但不限于：填充剂、稳定剂(例如海藻糖、甘氨酸)、矫味剂(例如木糖醇)、崩解剂(例如羧甲基纤维素钠)、粘合剂(例如明胶)、润滑剂(例如硬脂酸镁)。
在一些实施方案中，稳定剂选自以下的一种或组合：甘氨酸、赖氨酸、精氨酸、羟乙基淀粉、羟甲基淀粉、海藻糖、葡聚糖。
在一些实施方案中，矫味剂选自以下的一种或组合：蔗糖、单糖、糖精钠、阿斯巴甜、山梨醇、木糖醇、甘露醇。
在一些实施方案中，粘合剂选自以下的一种或组合：羧甲基纤维素钠、羟丙甲纤维素、明胶。
在一些实施方案中，润滑剂选自以下的一种或组合：有滑石粉、硬脂酸镁、微粉硅胶。
在一些具体的实施方案中，适用于本公开的药学上可接受的载体还可以提及，例如但不限于：右旋糖酐、乳糖、微晶纤维素、海藻糖、甘氨酸、木糖醇、羧甲基纤维素钠、赤藓糖醇、明胶、硬脂酸镁、抛射剂、保湿剂、溶剂、增溶剂、乳化剂、抗氧化剂、pH调节剂、防腐剂。具体而言，非限制实例还包括：白凡士林、卡波姆、羟丙甲纤维素、甲基纤维素、羟甲基纤维素钠、壳聚糖、硫糖铝壳聚糖、聚乙烯吡咯烷酮、聚乙烯醇、玻璃酸钠、二甲醚、四氟乙烷、氢氟烷烃、甘油、丙二醇、去离子水、注射用水、蒸馏水、乙醇、十六醇、十八醇、对氨基苯甲酸、乙酰胺、异丙醇、吐温、聚氧乙基氢化蓖麻油、硬脂酸、单硬脂酸甘油酯、三聚甘油单硬脂酸酯、脂肪酸蔗糖酯、蔗糖酯、乙酸异丁酸蔗糖糖酯、山梨醇酐三硬脂酸酯、肉豆蔻酸异丙酯、胆固 醇、角鲨烯、角鲨烷、正丁醇、乙二醇、乙醇、丙二醇、聚甘油酯、亚硫酸盐、半胱氨酸、二叔丁基羟基甲苯、山梨酸钾、磷酸缓冲溶液、三乙醇胺、氢氧化钠、乙二胺、月桂胺、碳酸氢钠、盐酸、尼泊金类、硫柳汞、氯甲酚、三氯叔丁醇、苯甲酸及其钠盐。
第四方面，本公开提供一种在受试者中预防或治疗伴有高危型HPV感染的慢性宫颈炎的方法，包括步骤：向受试者施用治疗有效量的本公开的红色诺卡氏菌细胞壁骨架或药物组合物。
“施用”、“提供给”、“处理”当应用于动物、人、细胞、组织、器官或生物样品时，是指药物或医疗装置与动物、人、细胞、组织、器官或生物样品接触。
“治疗”意指给予受试者内用或外用药物(治疗剂、活性成分或组合物)(如，本公开的外泌体或药物组合物)或医疗装置，在被治疗的受试者(或群体)中缓解(减轻、延迟、改善、治愈)一种或多种疾病症状，以至于达到临床可测量的程度，其中所述的受试者已经患有、疑似患有或易感于一种或多种疾病或其症状。
有效缓解任何疾病症状的药物(治疗剂、活性成分或组合物)的量，称为治疗有效量。可根据多种因素变化，例如受试者的疾病状态、年龄和体重。应当理解，在缓解单个受试者的目标疾病或其症状时，药物(治疗剂、活性成分或组合物)可能无效，但是根据本领域已知的任何统计学检验方法(如Student T检验、卡方检验、依据Mann和Whitney的U检验)确定，药物(治疗剂、活性成分或组合物)在统计学意义上对目标疾病或其症状是有效的。
在一些具体的实施方案中，受试者是人以外的动物，例如用于农场动物、宠物、工作动物、观赏动物、生产动物、实验动物(如大鼠、小鼠、豚鼠、兔、犬、灵长类)。
在一些具体的实施方案中，受试者是人。在一些具体的实施方案中，受试者疑似患有、确诊患有、已经患有、或易感于目标疾病或其症状。
在一些实施方案中，一天施用1-3次，或一天施用一次，或二天施用一次。视患者病灶的面积及程度不同，采用不同的剂量。
在一些实施方案中，施用周期持续2天至6月，例如，1周、2周、 3周、4周、5周、6周、7周、8周、9周、10周、11周、12周、13周、14周、15周、16周、17周、18周或更长、以及前述任意两个数值之间的范围。
同一药物活性成分或不同药物活性成分可一次性施用，或者可分成许多更小的单位剂量，以一定时间间隔施用。应理解治疗的确切剂量、持续时间、间隔时间是所治疾病的函数，并且可使用动物或临床试验数据推断而确定。所述施用可包括单次施用，或者间隔适当时间间隔的两次或更多次施用。其中，相邻两次的施用可间隔30分钟、40分钟、50分钟、60分钟、2小时、3小时、4小时、5小时、6小时、7小时、8小时、9小时、10小时、12小时、14小时、16小时、18小时、20小时、22小时、24小时、一天半、2天、3天、4天、5天、6天、7天、8天、9天、10天、1周、2周、3周、4周、5周、6周、1个月、2个月、3个月、4个月、5个月、6个月、7个月、8个月、9个月、10个月、11个月或12个月。
“任选”意味着其随后所描述的事项可以发生，但不必须发生；需要视情况而定。例如，“任选地，进行分装”意味着允许对产品进行分装，但是不是必须进行分装。
“一个”、“一”、“单个”、“该”，如果没有明确说明，也包括复数形式。
当提及数值范围时(比如60μg至120μg)，这是一种简写方式，视为将范围内的每一个值被明确提及，包括小数和整数。
具体实施方式
以下结合实施例进一步描述本公开。但这些实施例并非限制着本公开的范围。当未注明具体条件时，按照常规条件、按照原料供应商所建议的条件操作。未注明具体来源的试剂，为市场购买的常规试剂。
实施例
实施例1.市售的红色诺卡氏菌细胞壁骨架
红色诺卡氏菌细胞壁骨架购自辽宁格瑞仕特生物制药有限公司，国药准字S20030009(每支中细胞壁骨架的固体含量应不少于60μg， 其中胞壁酸含量不低于1.0μg，糖含量不低于4.0μg；复溶体积为2.0ml)。
实施例2.红色诺卡氏菌细胞壁骨架的制备
国药准字S20030009细胞壁骨架的制备方法与以下步骤本质上无显著差异，但因为生产规模不同，可进行调整。
因而，作为一个替代，可以采用以下步骤制备细胞壁骨架：
1.按照公知的方法培养菌体，并收集。对细胞进行破碎(例如超声波破碎或高压均质机破碎)。也允许采用本领域任何适当的公知方法对菌体进行破碎。显微镜下检查破碎的情况，每个视野有形菌不得超过5个，检查若干(10至30个)视野均达到此标准为合格。
2.除核酸：将破碎上清液进行离心，获得的沉淀物中加入DNA酶和RNA酶，按照酶的供应商建议的操作去除核酸。
3.除蛋白质：沉淀物加入常见的蛋白酶(例如胰蛋白酶)，按照酶的供应商建议的操作去除蛋白质。
4.除脂质：沉淀物中加入有机试剂(如丙酮、乙醚、乙醇中的一种或组合)，按照本领域常规操作去除脂质。
5.除细胞膜：沉淀物中加入TritonX-100，离心收集沉淀物，用PBS漂洗。
应当理解，上述除去杂质的步骤之间，技术人员可以调整先后顺序，使得步骤之间兼容。去除非细胞壁成分后，将沉淀物复溶于注射用水，待用。任选地，可以在115℃下灭菌20-30分钟，作为细胞壁骨架的原液。
实施例3.药物组合物的示例性制备方法
1.将实施例1或2所得产物涂覆在敷料(例如无菌纱布)上，制备成外用药物。
2.将实施例2所得产物制成冻干粉。
3.也可以采用本领域公知的洗剂制备方法，例如：洗剂多以水和醇为分散介质；由活性成分、电解质、等渗调节剂等在分散介质中制成。
4.将实施例1或2所得产物制备成胶囊。
5.将实施例1或2所得产物溶于注射用水制成注射剂。
6.将实施例1或2所得产物制备成阴道栓。
效果例
向临床确诊为慢性宫颈炎的受试者，施用实施例1的细胞壁骨架。治疗结果见表1。
表1

*HPV(-)是指高危型HPV测试均成阴性。

Claims (4)

  1. 红色诺卡氏菌细胞壁骨架在制备药物中的用途，其中：
    所述药物用于在受试者中预防或治疗伴有高危型HPV感染的慢性宫颈炎；
    所述高危型HPV选自以下的任一项或其组合：16、18、31、33、35、39、45、51、52、53、56、58、59、66、68型；
    所述慢性宫颈炎选自以下的任一项或其组合：宫颈柱状上皮异位、宫颈息肉、宫颈粘膜炎、宫颈腺囊肿、宫颈肥大。
  2. 根据权利要求1所述的用途，所述药物制备成选自以下任一项的剂型：膏剂、霜剂、栓剂、乳液、混悬剂、糊剂、凝胶剂、洗剂、片剂、气雾剂、喷雾剂、搽剂、粉剂、敷料、绷带、膜、贴片、注射剂。
  3. 根据权利要求1或2所述的用途，其中所述红色诺卡氏菌细胞壁骨架是国药准字S20030009或其等同标准对应的红色诺卡氏菌细胞壁骨架。
  4. 根据权利要求1或2所述的用途，其中所述红色诺卡氏菌细胞壁骨架通过以下方法获得，所述方法包括以下步骤或由以下步骤组成：
    1)提供红色诺卡氏菌；
    2)破碎所述红色诺卡氏菌，得到破碎产物；
    3.1)对所述破碎产物去除脂质；
    3.2)对所述破碎产物去除核酸；
    3.3)对所述破碎产物去除蛋白质；
    3.4)对所述破碎产物去除细胞膜；
    3.5)得到红色诺卡氏菌细胞壁骨架；
    4)任选地，分装；
    5)任选地，对所述红色诺卡氏菌细胞壁骨架进行冷冻干燥；
    其中，
    步骤3.1)、3.2)、3.3)、3.4)能够互换或并行，
    步骤4)和步骤5)能够互换；
    所述破碎的平均粒度为10nm至1000nm，优选10nm至800nm，更优选10nm至500nm。
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