探讨抗生素发展的新动态

1、 探讨抗生素发展的新动态中国药典各部收载的药品和生物制品是国家为保证药品生物制品质量制订的具有约束力的技术法规。它不仅是强制性的法定标准，而且还将起到指导药品和生物制品生产，保证药品生物制品质量，促进对外贸易的重要作用，同时还作为药品上市后技术监督的法定依据，因此是一部集专业性、技术性、法规性于一体的法典。2010年7月1日开始实施中国药典2010年版，在新版药典中抗生素品种做了一些必要的增加和调整，对某些检查项目进行了相应的修订。Chinese Pharmacopoeia of each department of pharmaceutical and biological products

2、 used is the country in order to ensure the quality of pharmaceutical and biological products for binding technical regulations. It is not only the statutory standards mandatory, but will also play a guiding drugs and biological products production, ensure the quality of pharmaceutical and biologica

3、l products, an important role in promoting foreign trade, but also as the legal basis of post-marketing drug technical supervision, so it is a set of professional, technical, and in one of the code. July 1, 2010 Chinese Pharmacopoeia 2010 edition of implementation, in the new edition of the Pharmaco

4、poeia antibiotics do some necessary to increase and adjust, some inspection items for the corresponding revision.1.1 中国药典2010年版中抗生素收载品种的主要变化在中国药典2010年版中新增加的品种及制剂总共76个，其中喹诺酮类抗生素为16个，说明我国在喹诺酮类抗生素方面的研究比较迅速。喹诺酮类抗生素不同于其他微生物来源的抗生素，由于其源于化学全合成，因此对于在国外已上市，我国临床上需要、但又无专利约束的喹诺酮类抗生素，国内基本上都已有生产。 新版药典对中国药典2000年版2中

5、收载的183个抗生素品种及制剂中的132个品种的方法或项目进行了不同程度的修订。Varieties and preparation of antibiotic 1.1 main changes of Chinese Pharmacopoeia 2010 edition collection contains varieties of increase in Chinese Pharmacopoeia 2010 edition of a total of 76, of which the quinolones antibiotics was 16, that of the quinolone a

6、ntibiotics aspect in our country rapidly. Quinolones is different from other microbial antibiotics, due to its source in chemical synthesis, therefore have been listed abroad, need, our country clinical but no patent constraints of quinolone antibiotics, domestic basically have the production. Metho

7、ds 132 varieties and 183 antibiotics of Chinese Pharmacopoeia 2000 edition of Pharmacopoeia ( 2 ) collection contains and preparation of or items were revised in different degree.1.2 国内有生产但未收载入中国药典2010年版的品种情况一些国内有生产的抗生素品种及制剂，由于其稳定性以及复方制剂组方的合理性需要进一步的验证，所以暂时未收载入新版药典中。主要品种有：国外尚未上市的喹诺酮类抗生素的注射液；国内自行研制的、用

8、不同酸制成的注射用阿奇霉素；国内自行研制的各种内酰胺类抗生素与不同的内酰胺酶抑制剂组成的复方制剂；以及甲氧苄嘧啶(TMP)与非磺胺类的抗生素(如头孢氨苄、庆大霉素、四环素等)组成的复方制剂。1.2 domestic production but did not receive varieties in China Pharmacopoeia 2010 edition of some domestic varieties and preparation of antibiotic production, because of its stability and composition of co

9、mpound preparation of rationality needs further verification, so for the time being not loaded in the codex. The main varieties are: injection of quinolone antibiotics have not yet listed abroad; Azithromycin domestic self-developed, made by different acid injection; compound preparation composed of

10、 beta-lactamase inhibitor of domestic self-developed beta lactam antibiotics with differentwithin; and trimethoprim ( TMP ) and non sulfa antibiotics ( such as cephalexin, gentamicin, tetracycline ) compound preparation.1.3 在中国药典2010年版的具体抗生素品种增修订过程中涉及到的相关工作(1)立足中国实际的国情，充分考虑城市和农村在用药方面的不同点，在中国药典2010年版

11、的抗生素品种中新增了价格相对比较便宜的麦白霉素。麦白霉素系我国自行研制的以麦迪霉素和柱晶白霉素为主要成分的混合物，我国药检工作人员用国外麦迪霉素的标准品标定了我国麦白霉素的效价，使麦白霉素的抗菌活性与麦迪霉素等值，同时在原料的质量标准中对其组分和效价均予以控制，使制剂在使用过程中不仅疗效明确，而且质量可控，可以满足广大农村群众的用药需要。(2)根据我国抗生素产品的具体情况，对现行质量标准进行修订、完善。如吉他霉素，起源于日本，但是由于我国菌种与日本菌种存在差异，我国生产的吉他霉素的各组分含量与日本吉他霉素有所不同。在中国药典2010年版的吉他霉素的质量标准中，为有效地保证产品质量，在吉他霉素原

12、料和制剂的药典标准中对各组分的含量都进行了控制，另外由于大环内酯类抗生素本身溶解性相对较差，因此在制剂的检查项目中增加了溶出度的检查项，而在日本的药典中对吉他霉素制剂的组分和溶出度均无相应的控制。(3)在中国药典2010年版中删除了乳酸环丙沙星原料。由于长期以来，人们认识上的误区，认为如果有药品制剂，一定应有其原料，因而在中国药典2000年版中收载了乳酸环丙沙星碱原料。但是，经过资料查阅和实验证明，鉴于乳酸环丙沙星原料的晶型和化学性质不稳定，国外制备乳酸环丙沙星注射液的原料并非用乳酸环丙沙星，而是用乳酸加环丙沙星碱制成的。乳酸环丙沙星原料并没有存在的必要，因此在新版的药典中删除了该品种。(4)

13、在新药试行标准转正的过程中，积极听取厂家的反馈意见。例如，美洛西林钠的成盐工艺以前均采用冷冻干燥或喷雾干燥方法，为无定型产品，纯度较低，含量的下限设为870%，现有的生产单位改为溶媒结晶工艺，成品为结晶型产品，含量可以提高到910%以上，但两种工艺得到的产品的红外光谱图不尽相同。为鼓励先进的生产工艺，根据生产厂家的意见，拟对美洛西林品种项下的检查项目进行相应的修订。1.3 in the Chinese Pharmacopoeia 2010 edition of specific antibiotics amendment related work relates to a process (

14、1 ) based on China's actual conditions, fully consider the city and countryside at different points in the medication, in Chinese Pharmacopoeia 2010 edition of antibiotics added price relatively cheap meleumycin. Meleumycin Department of China's mixture to Maddie kanamycin and leucomycin as

15、the main component of the self-developed, standard Chinese test staff with foreign Maddie mycin calibration titer in Chinese wheat mildew, make meleumycin antibacterial activity and Maddie mycin equivalent, at the same time in the quality standards of raw materials in the composition and valence wer

16、e control, make clear not only the efficacy of preparations in the process of using, and controllable quality, can meet the needs of the rural masses of medication. ( 2 ) according to the specific situation in China of antibiotic products, revise, perfect the current quality standard. As of kitasamy

17、cin, originated in Japan, but due to the strain differences with the Japanese culture, each element of China's production of guitar mycophenolate content and Japanese kitasamycin differ. In Chinese Pharmacopoeia 2010 edition of guitar mold quality standard element, in order to effectively guaran

18、tee the quality of products, in the standard of Pharmacopoeia of kitasamycin raw material and preparation of the contents of the components were control, also due to the relatively poor macrolide antibiotics is dissolved, the preparation of inspection items increased the stripping of the examination

19、, and in Japan in the Pharmacopoeia of kitasamycin preparation component and solubility were no corresponding control. ( 3 ) in the Chinese Pharmacopoeia 2010 removed the raw material of ciprofloxacin. For a long time, people's misunderstanding, think if there is medicine, certain its raw materi

20、als, thus containing the ciprofloxacin lactate alkali raw materials in Chinese Pharmacopoeia 2000 edition. However, through access to information and experiments show that, in view of crystal structure and chemical properties of ciprofloxacin lactate raw material is not stable, the preparation of ci

21、profloxacin lactate injection material not with ciprofloxacin lactate, but with lactic acid plus ciprofloxacin base. Ciprofloxacin lactate material and no need for its existence, therefore in the new edition of Pharmacopoeia to delete this variety. ( 4 ) in the process of tentative standards of new

22、drugs of positive, positive feedback manufacturers. For example, salt forming process of mezlocillin sodium before using freeze drying or spray drying method, is amorphous products, lower purity, content of the lower limit is set to 870%, the existing production units to solvent crystallization proc

23、ess, the finished product is crystalline, content can be increased to more than 910%, but the infrared spectra of two processing products are not the same. In order to encourage the advanced production technology, according to the manufacturer's opinion, of mezlocillin varieties under the inspec

24、tion items were revised.1.4 中国药典2010年版中抗生素品种增修订项目的特点在中国药典2010年版中主要对有关物质检查项、异常毒性检查项、梨苗细菌内毒素检查项、含量测定方法等项目进行了相应的修订。(1)有关物质检查项 对单组分抗生素，除哌拉西林、妥布霉素、硫酸巴龙霉素、硫酸核糖霉素、硫酸西索米星等少数品种外，基本上均制订了有关物质检查项。头孢羟氨苄、头孢氨苄、克拉霉素、盐酸四环素等品种项下的有关物质检查方法均由原来的薄层色谱法修订为高效液相色谱法。在阿莫西林、阿莫西林钠、克拉维酸钾、头孢唑林钠、头孢克洛、头孢地尼等品种的有关物质检查项中，均采用高效液相色谱法的梯度洗

25、脱，且限度与国外水平接近。(2)取消异常毒性检查项 随着分析技术的不断发展，许多抗生素中杂质的化学结构和药理性质已经研究得相当清楚，对于原来采用异常毒性检查来控制抗生素质量的做法已不适应目前的新情况，塑料土工格栅另外当采用动物实验进行异常毒性检查时，实验结果极易受实验环境和动物本身的影响，同时本着保护动物的原则，在中国药典2010年版中对注射用抗生素，林可类抗生素、头孢曲松、盐酸去甲万古霉素、硫酸卷曲霉素、放线菌素D、以及除硫酸链霉素以外的氨基糖苷类抗生素和口服的利福平均已取消了异常毒性的检查项。(3)细菌内毒素法的扩大应用 在中国药典2010年版中，除头孢替唑钠、头孢噻吩钠、盐酸去甲万古霉素

26、、盐酸四环素、硫酸多黏菌素B等少数几个品种还采用家兔热原法外，其它注射用抗生素基本上均采用了细菌内毒素法检查污染的发热性物质。但是，由于细菌内毒素检查法存在一定的局限性，只能检测革兰阴性菌污染的发热性物质，对于革兰阳性菌、真菌以及其他微生物污染的发热性物质则不能检出，并且在细菌内毒素实验中使用的鲎试剂来源于濒临灭绝的保护动物鲎，再加上有些抗生素本身对鲎试剂具有增强或抑制作用，干扰检测结果的可靠性，因而积极开展目前国外已在进行的“体外热原检测法”(In vitro Pyrogen Test，IPT)的研究是药品检验、药品研究、药品生产部门重要课题。(4)抗生素含量测定方法的修订 在中国药典201

27、0年版中，抗生素的含量测定方法主要从两个方面进行了修订。一是对原来用微生物效价测定效价的2个半合成的大环内酯类抗生素克拉霉素和罗红霉素修订为高效液相色谱法；二是对去甲万古霉素由原来采用HPLC法进行纯度测定和用微生物效价法测定效价的两种方法进行了统一，统一为高效液相色谱法测定含量。1.4 Chinese Pharmacopoeia 2010 edition of antibiotics amendment order project characteristics in Chinese Pharmacopoeia 2010 Edition focuses on the project rela

28、ted substances test, abnormal toxicity test, the bacterial endotoxin test items, content determination method for the corresponding revision. ( 1 ) the related substances of single component of antibiotics, in addition to piperacillin, tobramycin, Barone sulfate kanamycin, Ribostamycin sulfate, sulf

29、uric acid by magnitude few varieties, basically all formulated the related substances item. Determination of the related substances in varieties under cefadroxil, cephalexin, clarithromycin, tetracycline hydrochloride, by thin layer chromatography revised the original for high performance liquid chr

30、omatography. In the determination of related substances in terms of amoxicillin, amoxicillin sodium and clavulanate potassium, sodium, cefazolin, cefaclor, cefdinir varieties, gradient HPLC elution, and the limitation and foreign levels close to. ( 2 ) abolition of abnormal toxicity check with the c

31、ontinuous development of technology, many antibiotics and impurities in the chemical structure and pharmacological properties has been studied fairly clearly, for the original use of abnormal toxicity test to control the quality of antibiotics has not adapt to the new situation at present, in additi

32、on to the use of animal experiment for abnormal toxicity test, the experimental results extremely easy to be influenced by the experimental environment and the animal itself, at the same time in the animal protection principle, using antibiotics for injection in Chinese Pharmacopoeia 2010 edition, L

33、in can antibiotics, ceftriaxone, norvancomycin hydrochloride, capreomycin sulfate, actinomycin D, and the addition of streptomycin sulfate outside the aminoglycoside antibiotics and oral rifampicin has cancelled check average abnormal toxicity. ( 3 ) the bacterial endotoxin method applied in Chinese

34、 Pharmacopoeia 2010 edition, in addition to Ceftezole sodium, ceftriaxone sodium, norvancomycin hydrochloride, tetracycline hydrochloride, polymyxin B sulfate and a few varieties by using rabbit pyrogen method, material in the heating of the other injection adopts the bacterial endotoxin contaminati

35、on antibiotic medium. However, due to limitations of the bacterial endotoxins test, pyrogenic contamination can only be detected Gram-negative bacteria pollution, heating materials for Gram-positive bacteria, fungi and other microorganisms pollution can not be detected, and the use of bacterial endo

36、toxin test in the limulus reagent derived from Limulus endangered animal protection, and then plus some antibiotics itself has the enhancement or inhibition of limulus reagent, reliability, interference detection results, and actively carry out abroad in vitro pyrogen test method " " ( In

37、vitro Pyrogen Test, IPT ) research is an important subject of pharmaceutical production department of drug testing, drug research. ( 4 ) method for the determination of antibiotics in revision in Chinese Pharmacopoeia 2010 edition, method for determination of the content of antibiotics from two majo

38、r aspects of the revised. One is the original potency determination by microbiological assay of 2.5 synthetic macrolides clarithromycin and roxithromycin as amended by high performance liquid chromatography; two of norvancomycin by original HPLC method was used on two methods for determination of po

39、tency determination by microbiological assay and purity are unified, unified as determined by high performance liquid chromatography.1.5 新技术、新方法、新科研成果在中国药典2010年版中抗生素质量标准中的应用主要有以下几个方面：(1)蒸发光散射器作为检测器的高效液相色谱法的广泛应用3，4 蒸发光散射检测器(ELSD)由于它的通用性、响应因子一致性和梯度洗脱相容性等特点，越来越多的被应用于没有紫外吸收的化合物的分析中，在中国药典2010年版中，采用HPLCEL

40、SD法控制仅有末端紫外吸收的氨基糖苷类抗生素的组分以及含量，具体品种有硫酸依替米星、奈替米星、阿米卡星、庆大霉素、卡那霉素等。(2)庆大霉素的效价改用浊度法测定 抗生素效价的生物测定方法主要有琼脂扩散法(或管碟法)和浊度法，各国药典均已收载。管蝶法的特点为结果较为稳定，基本操作和设计适用于各种抗生素，并适用于多批样品的测定，但由于样品和标准品必须同质，且受扩散因素的影响，另外试验时间长(第二天才能得到结果)，手工操作，影响因素多，误差较大；相比之下，浊度法具有快速、易操作、不受扩散因素的影响，且可用仪器自动测量，试验的灵敏度和精密度均优于管蝶法。随着国产仪器质量的提高，中国药典2010年版附录

41、首次收载了浊度法，在抗生素正文中，庆大霉素采用了浊度法测定效价，不仅解决了管碟法测定庆大霉素效价误差大的问题，而且为解决多组分抗生素效价测定的困难开拓了新的思路。(3)高分子杂质检查方法的扩大应用 内酰胺类抗生素中的高分子杂质是引发临床上过敏反应的主要过敏原，系一类高分子聚合物。在中国药典2000年版中，已经收载4个头孢菌素类抗生素的高聚物检查方法。随着内酰胺类抗生素中高聚物检查方法的不断改进和完善，专用检测仪器的研制成功和标准化，中国药典2010年版中已经扩大到对11个内酰胺类抗生素21个品种都对其中的高聚物进行了控制，不仅促进了这些抗生素质量提高而且也保证了它们在临床上使用的安全性，这是我

42、国药品检验的科研成果转化为生产力的具体表现，在国际上是一个创新。目前国内药检工作者还在对检测方法作进一步的改进和完善，尝试采用短柱进行分析，以达到缩短分析时间的效果。另外，国内青霉素主要生产厂家正在研究低敏青霉素的生产工艺，以减少临床上过敏反应的发生。(4)氯霉素原料中对硝基苯甲醛的检查 在氯霉素的质量研究中，发现氯霉素原料中除能检查出氯霉素二醇物外，还能同时检测出对硝基苯甲醛，故在中国药典2010年版中增加了氯霉素中对硝基苯甲醛的限度规定，采用高效液相色谱法进行测定，限度不得过05%，此项检查在国外药典中尚未见有收载。(5)舒巴坦钠中特征性杂质控制方法的改进 在中国药典2010年版中，舒巴坦

43、钠中舒巴坦青霉胺杂质的检查法在国外主成分自身对照法的基础上修订为校正因子法(F=03)，缩短了分析时间。(6)固体口服制剂溶出度检查仍继续采用自身对照法 由于自身对照法具有消除样品中辅料干扰和不需要对照品的优点，中国药典2010年版中，头孢羟氨苄、头孢呋辛酯、头孢克洛、大环内酯类抗生素以及氨基糖苷类抗生素的口服固体制剂溶出度检查项均采用自身对照法测定。1.5 new scientific research of new technology, new method, used in Chinese Pharmacopoeia 2010 edition of antibiotics in the

44、 quality standards are mainly in the following aspects: ( 1 ) widely used ( evaporative light scattering detector for high performance liquid chromatography 3, 4 ) with evaporative light scattering detector ( ELSD ) because of its versatility, the response factor consistency and gradient elution fea

45、tures such as compatibility, more and more is applied to the analysis of compounds without ultraviolet absorption in, in Chinese Pharmacopoeia 2010 edition, aminoglycoside antibiotics to control only at the end of the ultraviolet absorption by HPLCELSD method, the component and content, the specific

46、 varieties of etimicin sulfate, netilmicin, Amikacin, gentamicin, kanamycin. ( 2 ) determination of antibiotic titer potency of the gentamicin using turbidimetry bioassay methods mainly include agar diffusion method ( or tube plate method ) and turbidimetric method, have been recorded in Pharmacopoe

47、ia of each country. Characteristics of pipe butterfly method is more stable result, basic operation and design is applicable to a variety of antibiotics, and suitable for the determination of the samples, but due to the sample and standard must be homogeneous, and affected by the diffusion factors,

48、in addition to a long experimental time ( the very next day to get results ), manual operation, many influence factors, error by contrast, with larger; effects of fast, easy operation, not by the diffusion of turbidity, and available instrument automatic measurement, test sensitivity and precision a

49、re better than that of tube plate method. With the improvement of the quality of domestic instrument, the appendix of China Pharmacopoeia 2010 was first recorded in the turbidity method, antibiotic, gentamicin by turbidimetric method for the determination of potency, not only solve the problem of bi

50、g error in the determination of gentamicin titer of cup-plate method, but also to solve multicomponent antibiotics titer determination difficult opened up new ideas. ( 3 ) the expansion of polymer impurity inspection method of high molecular weight impurities application of beta-lactam antibiotics i

51、s the main allergens trigger clinical allergic reactions, is a class of polymers. In the Chinese Pharmacopoeia 2000 edition, polymer inspection method has recorded 4 cephalosporins. With the development of polymer inspection method of beta-lactam antibiotics in the unceasing improvement and the cons

52、ummation, the successful development and standardization of the special detecting instrument, Chinese Pharmacopoeia 2010 edition has been expanded to 11 beta lactam antibiotics of 21 varieties of the polymer to control, not only to promote and improve the quality of these antibiotics but also to ens

53、ure the safety of their use in clinic, which is China's pharmaceutical inspection research into specific performance of productivity, in on international is an innovation. The current domestic testing workers are on the detection method for further improvement and perfection, try using a short c

54、olumn are analyzed, in order to shorten the analysis time effect. In addition, the production process of domestic penicillin major manufacturers are of low sensitivity to penicillin, to reduce the incidence of clinical allergy reaction. ( 4 ) chloramphenicol raw material p-nitrobenzaldehyde check on

55、 quality of chloramphenicol in chloramphenicol, found in raw material in addition to check out the glycols, can simultaneously detect p-nitrobenzaldehyde, so in the Chinese Pharmacopoeia 2010 increased the limit of p-nitrobenzaldehyde, chloramphenicol, by high performance liquid chromatography deter

56、mination, limit shall not exceed 5%, the check is not contained in foreign pharmacopoeia. ( 5 ) control method of characteristic of impurity sulbactam improved in Chinese Pharmacopoeia 2010 edition, basic examination Shubatan penicillamine impurity sulbactam in foreign main component self-compare on

57、 revisions to the correction factor method ( F=03 ), short analysis time. ( 6 ) check to continue using their control because of its control method can eliminate the interference of excipients in sample and advantage of not requiring control of release solid oral dosage forms, Chinese Pharmacopoeia

58、2010 edition, inspection items are using their own control method for the determination of dissolved solid oral preparation of cefadroxil, cefuroxime, cefaclor, big macrolide antibiotics, aminoglycoside antibiotics.2 国外关于新药质量研究的新动态2 foreign research on drug quality trends人用药品注册技术要求国际协调会(ICH)系由美国、日本和

59、欧盟三方政府药品管理部门和制药企业协会于1990年共同发起，旨在协调各国对注册药品所需的质量(Q)、安全(S)和疗效(E)三方面所需的技术资料，自1991年到2003年共举行了6次会议，在药品质量控制方面则从药品稳定性、方法学验证、杂质、质量标准等7个方面共制定了19个指导原则，在2003年第六次ICH会议以后，在质量部分又增加了四个指导原则，分别为Q8药物研发(Pharmaceutical Development);Q9质量风险管理(Quality Risk Analysis);Q4B药品管理部门对药典的互认(Regulatory Acceptance of Pharmacopoeia In

60、terchangeability)和Q5E生产工艺变动对生物技术产品/生物制品的可比性(Comparability of Biotechnology/Biological Products Subject to Changes in their Manufacturing Process)。药物研发是2004年11月由ICH指导委员会公布供ICH三方(美、欧、日)的药品管理部门协商讨论的关于药品质量的一个新指导原则，其主要系对原料的理化、生物学性质、辅料的功能特性、处方组成、生产工艺、包装材料、相容性、终产品稳定性等新药研究过程中获得的技术数据及灰质量控制要点，均作为过程控制和成品质量标准的基础，