CN103272134B - 一种杀灭抗酸染色阳性(红色)杆菌的药物 - Google Patents
一种杀灭抗酸染色阳性(红色)杆菌的药物 Download PDFInfo
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Abstract
本发明的目的在于提供一种杀灭抗酸染色阳性(红色)杆菌的药物,其特征在于:所述药物以碱性品红为活性成分,能够有效杀灭抗酸染色阳性杆菌。该药物由于使用无毒的碱性品红为活性成分,药物无毒副作用,患者服用后无不良反应,且取材方便,有利于推广应用,其作用机理与现有药物不同,能够彻底清除人体内的结核杆菌以及麻风杆菌,从而治愈结核病以及麻风病。
Description
技术领域
本发明属于医药技术领域,具体提供一种以检验和其它化工用途的原料——碱性品红为主要活性成分,以传统的中药和食品为辅料加工而成的根治结核和麻风病的药物——杀灭抗酸染色阳性(红色)杆菌的药物。
背景技术
结核病是由结核分枝杆菌引起的疾病。结核杆菌是一种位于细胞内感染的病原体,它能适应酸性环境,同时可长时间对抗巨噬细胞的吞噬体,而避免环境的酸化,使细菌在体内长时间的生存,所以结核病是一种持久的、慢性感染性疾病。结核菌经呼吸道进入体内,经过血液循环可寄生在全身各个器官进行繁殖,甚至可以进入骨、淋巴结中。目前最常见的是肺结核,它通过空气传播,危害大,流传广,有隐蔽性,带菌者无明显特征,防范困难,是人类的一大难题,长时间危害人体健康。
由于耐药菌株的出现和免疫抑制剂的应用,使全球范围内结核病的疫情骤然恶化,据WHO统计,全球约每三个人中就有一个人感染了结核分枝杆菌,在某些发展中国家,成人中结核分枝杆菌携带率高达80%,其中约有5—8%携带者可发展为活动性结核病。
目前全世界每年约出现八百万结核的新病例,并导致约三百万人死亡,中国每年死于结核病的人约有二十五万之多,是各类传染病死亡人数总和的二倍多。因此结核病是危害人体健康的全球性的问题。
结核菌通过呼吸道侵入人体,细菌在肺胞巨噬细胞内存活和复制,进而扩散至邻近非活化的肺泡并形成早期的感染灶,在免疫细胞反映期,结核菌在巨噬细胞内的最初生长,形成中心呈固态干酪坏死的结核灶,它能限制结核杆菌继续复制。对于大部分感染者来说,结核菌与宿主处于共生存的状态,结核菌被纤维包裹在干酪坏死的病灶内,此时结核菌不能增值。但一旦人体免疫损害,机体抵抗力下降,便可引起受抑制的结核菌重新的活动与繁殖,大量的结核菌从液化干酪灶释放出来播散到全身。对于这些原发和继发的病人要实施用药治疗。
目前常规的一线用药:1异烟肼
2.利福平
3.链霉素
4.吡嗪酰胺
5.乙胺丁醇
这些药都有很大的毒副作用(如异烟肼和利福平易引起血清谷丙转氨酶升高,肝肿大和黄疸、恶心、呕吐,链霉素损伤听觉神经,乙胺丁醇损害视神经等),且在治疗的过程中,这些药物没有使细菌死掉,而是在体内终身存留,只是抑制细菌不繁殖了,其治疗效果只是痰结核杆菌转阴(查不到抗酸杆菌),使细菌在体内又被包裹隐藏起来。一旦机体抵抗力弱或感冒,细菌又会在体内繁殖发病,造成很大痛苦,唯一的办法只有再实施抗痨治疗。由于药物的毒副作用能使患者的肝脏、肾脏、神经胃肠及造血器官受损,如谷草转氨酶、谷丙转氨酶、尿素、肌酐、尿素氮等极度异常,有的可出现血尿,医生只好将治疗结核的方案转移到保肝肾上来。很多患者结核病没治好而肝、肾等器官受了很大的伤害。
麻风病:麻风分枝杆菌是麻风病的病原体,此菌与其他分枝杆菌的不同点在于不能在人工培育基上生长,因此麻风的诊断仅依靠涂片做抗酸染色的检查。
麻风分枝杆菌和结核分枝杆菌的共同特点,都不易着色,一旦着色后,又不能为酸,酒精所褪色,故又称为抗酸杆菌。两者均为无鞭毛,无芽胞,无荚膜的革兰氏(G+)阳性杆菌。(经抗酸染色后都为红色(+))
麻风分枝杆菌和结核分枝杆菌在分类学上都属于裂殖菌纲,放线菌目,分枝杆菌科。麻风分枝杆菌和结核分枝杆菌区别如表1:
表1结核分枝杆菌和麻风分枝杆菌区别
目前治疗麻风病的用药与治疗结核病的药物基本相同,只是剂量更大、疗程更长,常用药有利福平、氯苯酚嗪、硫酰胺等,都是毒性极强的药物,对肝脏、肾脏、神经、胃肠、及造血功能都有严重的损害。
碱性品红(别称:碱性新品红、盐基品红、玫瑰苯胺盐酸盐):
1、碱性品红的基本用途:
碱性品红为最强的核染料,能使粘朊弹性组织和嗜品红的颗粒染色,中枢神经的核染色剂,细菌学用于鉴别结核杆菌,分析化学中配制席夫试剂检验醛类,用溴酸盐滴定的氧化还原指示剂,用于染棉、麻、人造纤维麦秆、皮革羽毛、脂肪等,也用于制造色淀。
2、制备来源:
由苯胺、邻甲苯胺,对甲苯胺的盐酸盐在铁和氯化锌存在下与硝基苯熔融缩合,再经酸提,中和结晶而制成。
3、化学结构式:
结构式说明:
用于细菌染色的染料,大部分是人工合成的含苯环的有机化合物(碱性品红是其中的一种),在其苯环上带有色基(chromophcre group)与助色基(auxochrome group)。硝基(—NO2)与偶氮基(—N=N—)等为色基,羟基(—OH)与氨基(—NH2)为助色基。色基能使该化合物具有颜色,含有色基的苯环化合物称为色原,这样的化合物不是染料,因它与被染物无亲和力,二者易分开,所以必须有助色基,助色基并不显颜色,但它本身能解离,给予化合物以离子特性。解离后的染料可以与被染物结合,使之着色。
助色基有的是碱性(如—NH2基);有的是酸性(如—OH基)。—NH2或—OH数目愈多,则碱性或酸性愈强。如每种基团各有一个时,则碱性占优势。因此可按照助色基的性质,来决定染料酸碱性。而碱性品红结构无—OH基,所以是碱性染料。
由于有机染料是带色的有机酸或碱类,难溶于水,而溶于有机溶剂,为使它们易溶解于水,通常均制成盐类。所以碱性品红也称盐基品红。综上所述,以上两个结构式只相差一个助色基(CH2),是一种物质在溶液中以两种结构的形式存在。
抗酸杆菌染色原理:
抗酸性杆菌含有分枝杆菌酸(mycolicacid),碱性品红(C20H20N3CL或C19H18N3CL)在类脂中的溶解度高于盐酸酒精,故两者能牢固结合,不易被盐酸酒精脱色。此外在染色过程中保持菌体表层结构的完整性,也是维持抗酸性的重要条件。因为完整的细胞壁,可阻止被碱性品红着色的类脂外溢。所以抗酸杆菌虽经脱色仍能保持红色。非抗酸杆菌不含分枝菌酸,不能抵抗盐酸酒精脱色,故可被美蓝复染成蓝色。
目前对细菌染色的机理尚未完全清楚,从已有的证据可以看出,染色过程既不是单纯的物理作用,也不是单纯的化学作用,而是两者的结合。
根据抗酸杆菌的特点,抗酸性染色法主要用于结核分枝杆菌和麻风分枝杆菌的检查。
4、分子式及分子量:
式Ⅰ:C20H20N3CL=337.84
式Ⅱ:C19H18N3CL=323.82
5、性状:为绿色金属光泽结晶,溶于乙醇和戊醇,微溶于水,在水溶液呈红色,不溶于乙醚。
发明内容
本发明的目的在于提供一种杀灭抗酸染色阳性(红色)杆菌的药物,该药物能够有效杀灭抗酸染色阳性(红色)杆菌,彻底清除人体内的结核杆菌以及麻风杆菌,从而治愈结核病以及麻风病。
本发明具体提供了一种杀灭抗酸染色阳性(红色)杆菌的药物,其特征在于:所述药物以碱性品红为活性成分。
本发明提供的杀灭抗酸染色阳性(红色)杆菌的药物,其特征在于:所述药物含有碱性品红药学上可接受的辅料或辅助性成分。其中药学上可接受的辅料或辅助性成分优选为米粞、小麦胚芽或五谷类辅料,最优选为米粞,选取米粞为辅料时,药物组成优选为重量百分比:碱性品红55-65%、米粞35-45%。
米粞的营养成分:富含有纤维素和大量的VB、VE族的维生素和矿物质,如钙、磷、铁等,俗称为肠道的清道夫,改善胃肠功能,增加肠的蠕动,清肠毒,及时将垃圾排出体外,解除便秘,促进营养的吸收,促进血液循环,提高免疫力,增强体质,米粞富含的膳食纤维还能与胆汁中的胆固醇结合,促进胆固醇的排出,有净化血液、调节血脂等作用。
小麦胚芽:含有高浓度的维持生命运动的各种营养成分,如脂肪、蛋白质及各种酵素、维生素、矿物质等,所含的维生素B1为每天人体需要的3.5倍。此外还含有大量的维生素E和罂酸,以及在其它食品中很少存在的维生素B6。
本发明提供的杀灭抗酸染色阳性(红色)杆菌的药物,其特征在于:所述的药物是口服制剂,注射制剂或外用制剂。
本发明提供的杀灭抗酸染色阳性(红色)杆菌的药物,其特征在于:所述药物是片剂、口服液、胶囊剂、注射剂、膏剂、酊剂、散剂、冲剂、栓剂、霜剂、透皮制剂或保健食品。
本发明还提供了碱性品红在制备抑制或/和杀灭抗酸染色阳性(红色)杆菌的药物以及在制备治疗结核病或/和麻风病的药物中的用途。
体内pH值的升高有利于清除结核杆菌和麻风杆菌:
人体最适的pH值在7.35-7.45之间,此时机体的免疫力最强,各种酶的活性也最高,体液是弱碱性环境,有利于新陈代谢。
当人体被结核菌感染时,因细菌的pH值约在2-5之间呈酸性,从而使人体pH值下降,呈酸性体质(也称酸败体质),使机体免疫力降低,因pH值可直接影响人体酶类的活性,使免疫应答受阻,又因结核菌体成分复杂,所含脂类与其菌的毒力有关,如硫脂有抑制巨噬细胞内溶酶体(lysosome)和吞噬体(phagosome)的融合,使细菌在体内无法清除。
当患者服用碱性品红时,体液(排出的尿液)的pH值有明显升高的趋势。而使被细菌感染的酸性体质恢复到弱碱性体质,有利于增强人体的免疫力,使结核菌被抑制或清除,因无法阻止巨噬细胞内溶酶体和吞噬体的融合,从而可清除细菌。服药后患者尿液pH值变化见表2和图1:
表2服药后患者(素食)在不同时间尿液pH值的变化
注:尿液pH值在服药后2-3小时达到高峰,且高峰期可持续3-4小时,并在服药后5-6小时恢复正常。
经我们的研究发现,碱性品红可以用于制备治疗结核病和麻风病的药物。且目前我们发现用碱性品红治疗结核病优于现有技术治疗结核病。其治疗优点如下:
(1)、无毒:用于口服两年的患者其肝、肾无损伤。
(2)、取材方便,有利于推广。
(3)、价格廉,给国家和个人减轻负担。
(4)、治疗彻底:治愈后的患者,无一例复发,体症消失一切结核病的症状。
(5)、无不良反应:在治疗过程中,无一例有副作用。
(6)、治疗方便:口服即可,不用住院。
(7)、药物不在体内存留,并使体液一时性有改善,服药后尿液的pH值变化如表2所示。
附图说明
图1服药后患者尿液pH值随时间变化曲线。
具体实施方式
本实施例所用碱性品红均购自天津钜华化工有限公司。
实施例1
碱性品红毒力实验:
材料:白色豚鼠共24只,购自沈阳药科大学,常规饲养;
碱性品红,天津钜华化工有限公司。
实验动物的配比分组:将实验用豚鼠随机分成4组(每组6只,雌雄各半),分别为空白对照组、碱性品红高、中、低剂量组。
给药方法:每只豚鼠每日上午固定时间给药一次,碱性品红高、中、低三个剂量组给药量分别为8mg/kg体重/d、6mg/kg体重/d、4mg/kg体重/d;给药时间为21天,每天观察3~4次,观察豚鼠精神、食欲、被毛光泽度、体格状况、警觉性以及躲避捕捉等能力。21天后未见任何变化。
实施例2
在体外给结核杆菌和麻风杆菌染色的原理上(见抗酸杆菌染色原理)可得知结核分枝杆菌和麻风分枝杆菌与碱性品红有特殊的亲和力,极易着色。
当服用碱性品红后,在不同的时间内碱性品红通过血液的现象可以看出,碱性品红以溶解的状态被吸收入血,而通过血细胞的过程,可以使吞噬细胞内的结核杆菌或麻风杆菌有着色或结合的机会,同时又提高了体液的pH值,破坏了细菌的生长环境,给细菌作了标记,又增强了免疫识别的能力,此时能促进机体强大的免疫系统充分发挥作用。因此碱性品红的碱性和细菌着色的作用是清除或杀灭细菌的所在。
血象观察:
一.观察对象:
1.健康者作正常人群观察对象
2.患者中痊愈的自愿者做实验对象
二.仪器:
双目生物显微镜xsp-12G型。
生产厂家:江光仪器总厂。
三.试剂:
1.瑞代-姬姆萨代染色液(市售)
2.磷酸盐缓冲液(市售)
四.操作方法:
1.取健康者和自愿者末梢血推片(正常对照血片,备用)。
2.取自愿者口服碱性品红后在不同时间的末梢血推片(口服碱性品红血片,备用)。
3.最后一次采血完成后,与未口服碱性品红之前的血片以及健康者末梢血推片(正常对照血片)一同染色。
五.在扩大100X的物镜(油镜)和扩大12.5X的目镜的显微镜中观察口服碱性品红血片,实物以扩大1250倍的状态下查看,结果见表3。
表3观察结果
观察结果:
1、服用碱性品红后,经人体吸收入血,通过细胞膜进入细胞内,在细胞内存留约2--4小时。
2、吞噬细胞内的结核杆菌或麻风杆菌有与碱性品红结合的机会,使其着色或消灭。
实施例3
药敏的检查培育:
(一)、药物:碱性品红
(二)、培养方法:罗—琴二氏培养基(改良法)
(三)、罗—琴二氏培养基的成分:
KH2PO4:2.4g、MgSO4·7H2O:0.24g、枸橼酸镁:0.6g、天门冬素:3.6g、甘油:12ml、蒸馏水:600ml、马铃薯粉:30g、新鲜全卵液:1000ml、2%孔雀绿水溶液:20ml。
(四)、仪器
1.电热恒温培育箱(黄石市恒丰医疗器械有限公司):36℃(特殊定制);
2.生物安全柜:BIOBASE BIOTEOH
规格型号:BSC—1500IZB2—Χ
生产厂家:济南鑫久西生物技术有限公司
3.蒸汽恒温箱:85℃
生产厂家:北京新中大业代表有限公司
4.药品:2%碱性品红水溶液,100ml备用。
(五)、培养基的制备:
将KH2PO4,MgSO4·7H2O,枸橼酸镁,天门冬素及甘油加水后,置于沸水浴中加热使之溶解,再加上马铃薯粉,继续在沸水中加热30分钟,不断搅拌,使其成均匀糊状。待冷却到60℃左右,加入新鲜全卵液1000ml,及2%孔雀绿水溶液20ml,充分混合后,用双层纱布过滤,分装于螺旋盖的试管中,每管加入7ml,此时将2%的碱性品红溶液按比例加入试管中(加入量见表3),将试管置于85%蒸汽恒温箱中90分钟,待冷至60℃以下时将试管倾斜放置在生物安全柜内(也称净化台),使培养基冷至凝固,并形成斜面,将菌液的浓度为10-2/ml的结核杆菌活菌液冲盈在培育基的斜面上。每管冲盈菌液为0.1ml。在培育基中菌液的浓度为10-3mg/ml,放入电热恒温培养箱中培养。培养五周后无菌生长为药物敏感,细菌仍然生长为不敏感(实验结果见表4)。
表4实验结果
细菌在药敏的培养基中生长的说明:
因为碱性品红无毒,在作药敏时,将碱性品红加入到罗—琴氏培养基中,只能使颜色有所变化,不会给培养基带来毒性,也不会改变罗—琴氏培养基的营养条件,又因培养基没有体细胞的环境,所以细菌应正常在此培养基中生长。更证明了碱性品红的无毒性,也证明了不是碱性品红本身具有杀灭抗酸染色阳性(红色)杆菌的作用,而是因为碱性品红的碱性使体液pH值升高,又给细菌菌体着上了颜色成为靶心,使人体内强大的免疫功能被激活,来彻底清除细菌,这是此药的极大优点。
如果在罗--琴氏培养基中加入常规药物,等于给细菌的食物下了毒,使细菌因中毒而不能繁殖,从而导致培养结果为细菌不能生长(称敏感)。即常规药物只能起到抑制细菌的作用,并不能彻底消灭细菌。且会带来严重的毒副作用,伤害人体的解毒器官。
碱性品红治疗结核杆菌和麻风杆菌的要点可概括为:
①、因碱性品红使机体的pH值升高,而使菌体失去了自身调节pH值的作用,无法抑制吞噬细胞分秘兴溶酶,使细菌被兴溶酶体和吞噬体的融合而清除。
②、因碱性品红给菌体着色,同时又提高了体液的pH值,即给细菌本身做了标记,又增强了免疫力,使吞噬细胞能识别目标,并增强吞噬的能力进而消灭细菌。
③、因碱性品红无毒,其作用机理与现有药物不同,无法用毒性抑制或杀灭细菌,所以在体外做药敏时因没有体细胞的环境,所以细菌不能被消灭,应该正常在培养基中生长。
实施例4
碱性品红治疗结核病:
患者用药量:胶囊500mg/粒,其中重量配比为碱性品红:米粞=3:2;每人口服1~2粒/日。
服用方法:每天早饭前服用碱性品红胶囊后即可用餐,三个月为一个疗程,患者病历见表5,治疗结果见表6。
病历说明:
1.此病历患者都是经过正规结核病医院或结核病防治所确诊的,且所有患者均经常规用药治疗后病情反复发作。
2.患者经正规治疗后有不同程度的肝脏、肾脏受损,而无法再服用常规药物。
3.在服药前都有明显的结核症状:周身无力,不适,发热,咳嗽,盗汗,腹水,吐血,呕血等(胸椎骨结核患者有疼痛,部分月经闭经)。
4.大多数患者服药一个月后症状明显减轻,三个月后消除结核症状,六个月经拍片确认痊愈。
5.服药后患者肝脏、肾脏抽血检查结果正常。
6.服药一个月后在痰中找不到结核肝菌(即未找到抗酸杆菌)。服药六个月后可以停药,能够恢复正常生活并参加体力劳动。
表5结核病患者个体病历表
表6结核病患者治疗效果
实施例5
碱性品红治疗麻风病:
一、药物种类:
1.口服药胶囊:患者用药量:胶囊500mg/粒,其中重量配比为碱性品红:米粞=3:2;每人口服1~2粒/日;
2.外用酊剂,涂抹溃疡面。
二、碱性品红酊剂的制备:
材料:1.碱性品红
2.医用酒精(75%乙醇溶液)
制备过程:
1.在玻璃烧杯内倒入100g碱性品红以及800ml75%的酒精,用玻璃棒搅拌均匀。
2.用镊子夹取酒精棉球,用打火机点燃后放在装有碱性品红和75%的酒精的烧杯表面,点燃混合液,燃烧时用玻璃棒搅拌,直至火焰燃尽。
3.待温度达到60℃时用漏斗将烧好的药液倒入磨口瓶中备用。
4.将制备好的酊剂浸入到有脱脂棉球的辅料缸中备用。
三.使用方法:
外用:先将麻风病人的溃疡面用盐水清洗干净,再用制备好的碱性品红酊剂脱脂棉球涂抹伤口,反复涂抹后用纱布遮盖包扎好,3天一换药,直至伤口愈合为止。
内服:每日早饭前用温开水口服1粒胶囊,直至痊愈,一个疗程为3个月,治疗效果见表7。
病历说明:对象为山东滕州市麻风村的麻风病患者以及广东省无川市普光医院麻风病康复中心的患者。
表7麻风病患者个体病历表
上述实施例只为说明本发明的技术构思及特点,其目的在于让熟悉此项技术的人士能够了解本发明的内容并据以实施,并不能以此限制本发明的保护范围。凡根据本发明精神实质所作的等效变化或修饰,都应涵盖在本发明的保护范围之内。
Claims (3)
1.一种杀灭抗酸染色阳性杆菌的药物,其特征在于:所述药物是片剂、口服液、胶囊剂、膏剂、酊剂、散剂、冲剂、栓剂或霜剂,以碱性品红为活性成分,还含有碱性品红药学上可接受的辅料或辅助性成分,所述辅料或辅助性成分为米粞。
2.按照权利要求1所述杀灭抗酸染色阳性杆菌的药物,其特征在于:所述药物其组成为重量百分比:碱性品红55-65%、米粞35-45%。
3.碱性品红在制备治疗麻风病的药物中的用途。
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