CN117337187A - 包含柠檬明串珠菌菌株作为有效成分的用于预防或治疗肠道损伤的药物组合物 - Google Patents
包含柠檬明串珠菌菌株作为有效成分的用于预防或治疗肠道损伤的药物组合物 Download PDFInfo
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- CN117337187A CN117337187A CN202280014024.5A CN202280014024A CN117337187A CN 117337187 A CN117337187 A CN 117337187A CN 202280014024 A CN202280014024 A CN 202280014024A CN 117337187 A CN117337187 A CN 117337187A
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Landscapes
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Abstract
本发明涉及一种用于预防、改善或治疗肠道损伤,具体地,炎症性肠病、非酒精性脂肪性肝炎及因化疗引起的肠道损伤的组合物,其包含柠檬明串珠菌菌株或其的培养物作为有效成分。根据本发明的组合物,对炎症性肠病、非酒精性脂肪性肝炎及因化疗等引起的各种肠道损伤表现出优异的预防和治疗效果,因此,可以有效地用作用于治疗、预防或改善人或动物的肠道损伤的组合物。
Description
技术领域
本发明涉及一种用于预防、改善或治疗肠道损伤(intestinal damage),具体地,炎症性肠病、非酒精性脂肪性肝炎及因化疗引起的肠道损伤的组合物,所述组合物包含柠檬明串珠菌(Leuconostoc citreum)菌株或其的培养物作为有效成分。
背景技术
现代人的以肉食为主的饮食习惯、以速食和加工食品为主的饮食习惯、过多摄入刺激性食物、因生活环境繁忙而急于摄取食物的饮食习惯等不良饮食习惯和不健康生活方式、以及随着年龄增长,有害菌战胜有益菌,肠道菌群失调,对肠道健康产生不利影响。
现代人暴露于各种肠道疾病,很多人不仅因炎症性肠病、非酒精性脂肪性肝炎、结肠癌等疾病而造成肠道损伤,而且,还因肠道损伤而导致治疗后维持肠道损伤恢复和功能等问题。
肠道菌群(microbiota)作为微生物菌群是指环境内的微生物群落,据报道,肠道菌群对宿主,例如人的免疫、代谢物质等的内环境稳定起到重要作用。肠道菌群与宿主交换化学物质信号,并且,基于肠道菌群的免疫细胞的表达或神经递质的产生、短链脂肪酸(Short chain fatty acids,SCFA)等对于宿主内的系统产生极大的影响,特别是益生菌/益生元可使宿主的不平衡的肠道菌群变成平衡状态,且这种健康的肠道菌群的代谢产物可促进宿主的健康。
目前,公开有包含益生菌作为有效成分的用于预防或治疗酒精性肠道损伤的组合物(韩国授权专利第10-2038695号)乃至包含新型嗜酸乳杆菌菌株的用于改善肠道环境的组合物(韩国授权专利第10-2201517),但尚未公开包含柠檬明串珠菌菌株作为有效成分的预防或治疗肠道损伤的效果。
在这种背景下,本发明人确认到包含柠檬明串珠菌菌株或其培养物的组合物对炎症性肠病、非酒精性脂肪性肝炎、因化疗等引起的受损肠道具有治疗肠道损伤及功能恢复的效果,以完成了本发明,所述组合物几乎没有毒性及副作用。
发明内容
本发明的目的在于,提供一种用于预防或治疗肠道损伤的药物组合物,其包含柠檬明串珠菌菌株作为有效成分。
另外,本发明的其他目的在于,提供一种用于预防或改善肠道损伤的食品组合物或食品添加剂组合物,其包含柠檬明串珠菌菌株作为有效成分。
另外,本发明的其他另一目的在于,提供一种用于预防或改善家畜的肠道损伤的饲料组合物或饲料添加剂组合物,其包含柠檬明串珠菌菌株作为有效成分。
为了达成所述目的,本发明提供一种用于预防或治疗肠道损伤的药物组合物,其包含柠檬明串珠菌菌株或其培养物作为有效成分。
另外,本发明提供一种用于预防或改善肠道损伤的食品组合物或食品添加剂组合物,其包含柠檬明串珠菌菌株或其培养物作为有效成分。
另外,本发明提供一种用于预防或改善家畜的肠道损伤的饲料组合物或饲料添加剂组合物,其包含柠檬明串珠菌菌株或其培养物作为有效成分。
发明效果
为了达成所述目的,本发明提供一种用于预防或治疗肠道损伤的药物组合物,其包含柠檬明串珠菌菌株或其培养物作为有效成分。
另外,本发明提供一种用于预防或改善肠道损伤的食品组合物或食品添加剂组合物,其包含柠檬明串珠菌菌株或其培养物作为有效成分。
另外,本发明提供一种用于预防或改善家畜的肠道损伤的饲料组合物或饲料添加剂组合物,其包含柠檬明串珠菌菌株或其培养物作为有效成分。
附图说明
图1是示出用根据本发明的组合物处理Caco2肠细胞后测量ZO1蛋白的基因表达量的图表的图。
图2是示出在CDHFD小鼠模型中处理根据本发明的组合物后测量Occludin蛋白的基因表达量的图表的图。
图3是示出在CDHFD小鼠模型中处理根据本发明的组合物后测量ZO1蛋白的基因表达量的图表的图。
具体实施方式
本发明的包含柠檬明串珠菌菌株或其培养物作为有效成分的组合物具有预防或治疗肠道损伤的效果,并且可以用作药物组合物。
本发明的柠檬明串珠菌菌株是乳酸菌菌株。所述柠檬明串珠菌菌株是一种益生菌,具有乳酸菌的一般的缓解肠道紊乱效果及免疫增强效果。众所周知,明串珠菌属的乳酸菌具有缓解肠道紊乱效果及免疫增强效果。
所述柠檬明串珠菌菌株,可以为柠檬明串珠菌WiKim0104菌株,并具有SEQ ID NO:1的核酸序列,但不限于此。
所述柠檬明串珠菌菌株可以在MRS液体培养基接种0.1%至10%,并在25℃至37℃的温度下培养4小时至48小时来使用。
所述培养方法优选为静态培养法,但不限于此。
在本发明中,“益生菌(probiotics)”是存活于包括人在内的动物的胃肠器官内,改善宿主的肠内微生物环境,从而对宿主的健康有利的微生物。益生菌是一种具有益生菌活性的活的微生物,其具有单一或复合菌株形态,当以干燥的细胞形态或发酵产物形态用于人或动物的情况下,能够对宿主的肠道菌群产生有利的影响。
所述肠道损伤可以为炎症性肠病、非酒精性脂肪性肝炎或化疗引起的肠道损伤,但不限于此。
包含在根据本发明的组合物中的柠檬明串珠菌菌株可以以活菌体或死菌体存在,还可以以干燥或冻干形式存在。另外,柠檬明串珠菌菌株的培养物可以为有效成分,并且在所述培养物可以包括活菌培养液或死菌上清液。适合包含在各种组合物中的乳酸菌的形态和配制方法是本领域技术人员众所周知的。
所述组合物可以通过口服或胃肠外给药。胃肠外给药可以通过静脉注射、皮下注射、肌肉注射、腹腔注射、经皮给药、局部给药、鼻内给药、肺内给药及直肠内给药等方式给药,优选地,可以通过静脉注射的方法给药,但不限定于此。
所述组合物的适当给药量可以根据配制方法、给药方式、患者的年龄、体重、性别、疾病状态、饮食、给药时间、给药途径、排泄速度及反应敏感性之类的因素给出多种处方。
在将本发明的组合物用作药物组合物的情况下,除上述有效成分外,本发明的药剂学组合物可以使用药剂学上有效、生理学上可接受的辅助剂来制备,所述辅助剂可以使用赋形剂、崩解剂、甜味剂、结合剂、包衣剂、膨胀剂、润滑剂、滑泽剂或香味剂等。
为了给药,除上述有效成分以外,所述药物组合物还可以通过包含一种以上的药学上可接受的载体来优选地配制为药物组合物。
例如,为了配制为片剂或胶囊剂,有效成分可以与乙醇、甘油、水等之类的口服、无毒性的药学上可接受的非活性载体结合。并且,可以根据需要包含适当的结合剂、润滑剂、崩解剂及发色剂或混合物。适当的结合剂包括淀粉、明胶、葡萄糖或β-乳糖之类的天然糖、玉米甜味剂、阿拉伯胶、黄芪胶或油酸钠之类的天然或合成胶、硬脂酸钠、硬脂酸镁、苯甲酸钠、醋酸钠、氯化钠等,但不限定于此。崩解剂包括淀粉、甲基纤维素、琼脂、膨润土、黄原胶等,但不限定于此。在以液体溶液的形态配制的组合物中可接受的药剂学载体可以使用适于灭菌及活体的生理盐水、灭菌水、林格氏液、缓冲生理盐水、白蛋白注射溶液、葡萄糖溶液、麦芽糊精溶液、甘油、乙醇及混合一种以上这些成分来使用,可以根据需要添加抗氧化剂、缓冲液、抑菌剂等其他常规的添加剂。并且,还可以添加稀释剂、分散剂、表面活性剂、结合剂及润滑剂来配制为水溶液、悬浮液、乳浊液等之类的注射用剂型、丸药、胶囊、颗粒或片剂。
进而,可以将Remington’s Pharmaceutical Science,Mack PublishingCompany,Easton PA中公开的方法用作本发明相关技术领域的适当的方法,根据各疾病或成分来优选地配制。
本发明的包含柠檬明串珠菌菌株或其培养物作为有效成分的组合物,可以用作用于预防或改善肠道损伤的食品组合物或食品添加剂组合物。
所述食品组合物可以以保健功能食品的形式存在。
所述“保健功能食品”是指根据有关保健功能食品的法律,使用具有对人体有用的功能性的原料或成分制备及加工的食品(第三条第一项),“功能性”是指通过调节营养素或生理学作用等之类的保健用途来在人体的结构及功能中获得有用的效果(同一条的第二项)。
所述食品组合物还可以包括食品添加物,若无特别规定,“食品添加物”是否适当则根据韩国食品药品监督管理局认可的韩国食品添加物法典的总则及通常的试验方法等并按照与相关品种有关的规格及标准来判定。
收录于上述“食品添加物法典”中的品种有:例如酮类、甘氨酸、柠檬酸钾、烟酸、肉桂酸等化学合成品、柿子色素、甘草提取物、结晶纤维素、瓜尔豆胶等天然添加物、L-谷氨酸钠制剂、面类添加碱剂、防腐剂、焦油色素制剂等混合制剂类。
包含本发明的有效成分的食品可以为面包、糕类、干果类、糖果类、巧克力类、口香糖、果酱类之类的饼干类、冰淇淋类、冰果类、冰淇淋粉末类之类的冰淇淋制品类、牛奶类、低脂牛奶类、乳糖分解牛奶、加工乳类、山羊奶、发酵乳类、黄油乳类、浓缩乳类、奶油类、黄油类、天然奶酪、加工奶酪、奶粉类、乳清类之类的乳加工品类、肉食加工品、蛋加工品、汉堡包之类的肉食制品类、鱼饼、火腿、香肠、培根等鱼肉制品之类的鱼肉制品类、拉面类、干面类、生面类、汤面类、豪华干面类、改良熟面类、冷冻面类、意大利面类之类的面类、水果饮料、蔬菜类饮料、碳酸饮料、豆乳类、酸奶等的乳酸菌饮料、混合饮料之类的饮料、酱油、黄豆酱、辣椒酱、干黄酱、清国酱、混合酱、食醋、酱类、番茄酱、咖喱、调味汁之类的调味食品、人造黄油、起酥油以及比萨,但不限定于此。
除上述以外,本发明的组合物还可以包含多种营养剂、维生素、电解质、风味剂、着色剂、果胶酸及其盐、海藻酸及其盐、有机酸、保护性胶体增稠剂、pH调节剂、稳定剂、防腐剂、甘油、乙醇、用于碳酸饮料的碳酸化剂等。此外,本发明的组合物还可以包含用来制备天然果汁、果汁饮料及蔬菜饮料的果肉。这些成分可单独或组合来使用。
包含本发明的有效成分的饮料组合物对其他成分没有特别限制,可以与普通饮料一样包含多种香味剂或天然碳水化合物等。上述天然碳水化合物的例有单糖(例如葡萄糖、果糖等)、二糖(例如麦芽糖、蔗糖等)、多糖(例如糊精、环糊精等)之类的普通的糖、以及木糖醇、山梨糖醇、赤藓糖醇等糖醇。除上述以外,还可以有利地将天然香味剂(索马甜、甜叶菊提取物(例如莱鲍迪甙A、甘草酸等))以及合成香味剂(糖精、阿巴斯甜等)用作香味剂。
并且,本发明的包含柠檬明串珠菌菌株或其培养物作为有效成分的组合物,可以用作用于预防或改善家畜的肠道损伤的饲料组合物或饲料添加剂组合物。
在将所述组合物制备为饲料添加剂的情况下,所述组合物可以制备为20%至90%的高浓缩液或粉末或颗粒形态。所述饲料添加剂还可以包含柠檬酸、富马酸、己二酸、乳酸、苹果酸等有机酸或磷酸钠、磷酸钾、酸性焦磷酸盐、聚磷酸盐(聚合磷酸盐)等磷酸盐或多酚、儿茶素、α生育酚、迷迭香提取物、维生素C、绿茶提取物、甘草提取物、甲壳素、鞣酸、植酸等天然抗氧化剂中的一种或一种以上。在制备为饲料的情况下,所述组合物可以配制为普通的饲料形态,可以包含普通的饲料成分。
所述饲料及饲料添加剂还可以包含谷物,例如由粉碎或破碎的小麦、燕麦、大麦、玉米及大米;植物蛋白饲料,例如以芸苔、大豆以及葵花为主要成分的饲料;动物蛋白饲料,例如血粉、肉粉、骨粉及鱼粉;以及糖分及乳制品,例如各种奶粉及乳浆粉末组成的干燥成分等,此外还可以包含营养补充剂、消化及吸收促进剂、生长促进剂等。
所述饲料添加剂可以单独向动物给药,或者在食用载体中与其他饲料添加剂组合来给药。并且,所述饲料添加剂可以在追肥饲料中,或是将其与动物饲料直接混合,或者在饲料外以单独剂型的方式轻松地向动物给药。在将所述饲料添加剂与动物饲料单独给药的情况下,可以与相关领域公知的药学上可接受的食用载体组合制备为即时释放或缓释剂型。所述食用载体可以为固体或液体载体,例如玉米淀粉、乳糖、蔗糖、玉米片、花生油、橄榄油、芝麻油及丙二醇。在使用固体载体的情况下,饲料添加剂可以为片剂、胶囊剂、散剂、含片或糖衣片剂或微分散性形态的追肥饲料。在使用液体载体的情况下,饲料添加剂可以为明胶软质胶囊剂,或者糖浆剂或悬浮液、乳液剂或溶液剂的剂型。
并且,所述饲料及饲料添加剂可以包含辅助剂,例如,保存剂、稳定剂、湿润剂或润滑剂、溶液促进剂等。所述饲料添加剂通过浸注、喷雾或混合的方式添加于动物的饲料来使用。
本发明的饲料或饲料添加剂可以适用于包括哺乳类、家禽及鱼类的大多数动物的饮食中。
作为所述哺乳类,不仅可以用于猪、牛、马、羊、兔子、啮齿类动物以及用于实验的啮齿动物即小鼠、仓鼠、豚鼠,还可以用于宠物(例如狗、猫)等,作为所述家禽类,可以用于鸡、火鸡、鸭、鹅、雉鸡以及鹌鹑等,作为所述鱼类,可以用于为鲤鱼及鳟鱼等,但不限定于此。
实施方式
下面,通过实施例对本发明进行更详细地说明。这些实施例仅用于更详细地说明本发明,并且,对于本领域的普通技术人员来说根据本发明的主旨本发明的范围不受这些实施例的限制是显而易见的。
实施例1.培养菌株
柠檬明串珠菌WiKim0104(保藏号KCCM12420P)菌株经保藏者韩国食品研究所许可分配得到,并进行了实验。
将所述分配得到的柠檬明串珠菌WiKim0104菌株以1%接种于30ml的MRS液体培养基中,在30℃的温度下静置培养18小时。培养后,以3000rpm的转速离心分离10分钟,然后单独保存培养液,并用磷酸盐缓冲溶液(PBS,phosphate buffered saline)洗涤菌体3次,以除去剩下的培养基成分。
实施例2.确认肠细胞中与屏障维持相关的基因表达增强功效
为了培养肠细胞(HT29细胞和Caco2),使用添加青霉素/链霉素和10%FBS的RPMI培养基,并使用6孔板(6-well)transwell进行做好准备。
培养肠细胞直至其生长汇合后,用200μl的含有1μg/ml的LPS和500μM的棕榈酸(Palmitic acid)的新培养基替换。此时,以1×108CFU/mL的浓度处理准备好的柠檬明串珠菌WiKim0104菌株。经所述菌株处理后,培养24小时,然后用PBS洗涤3次,收获细胞,并使用TRIZOL试剂分离RNA。将1μg的RNA使用cDNA合成试剂盒合成cDNA之后,通过qRT-PCR(quantitative real time polymerase chain reaction)方法确认紧密连接(Tightjunction)相关基因ZO-1的表达量,并将结果示于图1。
如图1所示,显示在柠檬明串珠菌WiKim0104菌株处理组中,由LPS和棕榈酸减少的ZO1的表达量得到恢复的结果。
实施例3.确认在非酒精性脂肪性肝炎模型中屏障维持功效
已知非酒精性脂肪性肝炎也与紧密连接(Tight junction)的功能障碍有密切相关。因此,在通过choline deficient high fat diet(CDHFD)方法诱导的非酒精性脂肪性肝炎模型中处理根据本发明的组合物之后,测量与紧密连接相关的基因的表达量。通过CDHFD方法对6-8周龄的C57BL6小鼠摄入8周后诱导非酒精性脂肪性肝炎,然后,以2×109CFU/天的浓度每周饲喂5天柠檬酸明串珠菌WiKim0104菌株,持续12周。12周后,确认对照组和实验组肠组织中紧密连接(Tight junction)相关基因Occludin和ZO1的表达量,将结果示于图2及图3。
如图2及3所示,确认到在摄取柠檬明串珠菌Wikim0104菌株的组中,所述Occludin和ZO1蛋白基因的表达量恢复至正常组的水平。
[保藏编号]
保藏机构名称:韩国微生物保藏中心(国外)
保藏编号:KCCM12420P
保藏日期:20181214
国际承认用于专利程序的微生物保藏
布达佩斯条约
国际格式
To.韩国食品研究院 本页底部指明的国际保藏机构
大韩民国全罗北道完州郡伊西面 据规则7.1出具的原件的收据
农生命路245,55356
在适用规则6.4(d)的情况下,该日期是获得国际保藏机构资格的日期。
序列表
<110> 利斯科生物科技(Liscure Biosciences)
<120> 柠檬明串珠菌 WiKim0104
<130> WiKim0104
<150> KR 10-2021-0017813
<151> 2021-02-08
<160> 1
<170> KoPatentIn 3.0
<210> 1
<211> 1430
<212> DNA
<213> 柠檬明串珠菌
<400> 1
gatgaacgct ggcggcgtgc ctaatacatg caagtcgaac gcgcagcgag aggtgcttgc 60
acctttcaag cgagtggcga acgggtgagt aacacgtgga taacctgcct caaggctggg 120
gataacattt ggaaacagat gctaataccg aataaaactt agtatcgcat gatatcaagt 180
taaaaggcgc tacggcgtca cctagagatg gatccgcggt gcattagtta gttggtgggg 240
taaaggctta ccaagacgat gatgcatagc cgagttgaga gactgatcgg ccacattggg 300
actgagacac ggcccaaact cctacgggag gctgcagtag ggaatcttcc acaatgggcg 360
caagcctgat ggagcaacgc cgcgtgtgtg atgaaggctt tcgggtcgta aagcactgtt 420
gtatgggaag aaatgctaaa atagggaatg attttagttt gacggtacca taccagaaag 480
ggacggctaa atacgtgcca gcagccgcgg taatacgtat gtcccgagcg ttatccggat 540
ttattgggcg taaagcgagc gcagacggtt gattaagtct gatgtgaaag cccggagctc 600
aactccggaa tggcattgga aactggttaa cttgagtgtt gtagaggtaa gtggaactcc 660
atgtgtagcg gtggaatgcg tagatatatg gaagaacacc agtggcgaag gcggcttact 720
ggacaacaac tgacgttgag gctcgaaagt gtgggtagca aacaggatta gataccctgg 780
tagtccacac cgtaaacgat gaatactagg tgttaggagg tttccgcctc ttagtgccga 840
agctaacgca ttaagtattc cgcctgggga gtacgaccgc aaggttgaaa ctcaaaggaa 900
ttgacgggga cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac 960
cttaccaggt cttgacatcc tttgaagctt ttagagatag aagtgttctc ttcggagaca 1020
aagtgacagg tggtgcatgg tcgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc 1080
gcaacgagcg caacccttat tgttagttgc cagcattcag ttgggcactc tagcgagact 1140
gccggtgaca aaccggagga aggcggggac gacgtcagat catcatgccc cttatgacct 1200
gggctacaca cgtgctacaa tggcgtatac aacgagttgc caacctgcga aggtgagcta 1260
atctcttaaa gtacgtctca gttcggactg cagtctgcaa ctcgactgca cgaagtcgga 1320
atcgctagta atcgcggatc agcacgccgc ggtgaatacg ttcccgggtc ttgtacacac 1380
cgcccgtcac accatgggag tttgtaatgc ccaaagccgg tggcctaacc 1430
Claims (12)
1.一种用于预防或治疗肠道损伤的药物组合物,其中,包含柠檬明串珠菌菌株或其培养物作为有效成分。
2.根据权利要求1所述的用于预防或治疗肠道损伤的药物组合物,其中,所述柠檬明串珠菌菌株是具有SEQ ID NO:1的核酸序列的柠檬明串珠菌WiKim0104(保藏编号KCCM12420P)菌株。
3.根据权利要求1所述的用于预防或治疗肠道损伤的药物组合物,其中,所述肠道损伤为炎症性肠病、非酒精性脂肪性肝炎及因化疗引起的肠道损伤。
4.根据权利要求1所述的用于预防或治疗肠道损伤的药物组合物,其中,所述组合物可以通过口服给药或胃肠外给药方式给药。
5.根据权利要求4所述的用于预防或治疗肠道损伤的药物组合物,其中,所述胃肠外给药可以通过静脉注射、皮下注射、肌肉注射、腹腔注射、经皮给药、局部给药、鼻内给药、肺内给药或直肠内给药。
6.一种用于预防或改善肠道损伤的食品组合物,其中,包含柠檬明串珠菌菌株或其培养物作为有效成分。
7.根据权利要求6所述的用于预防或改善肠道损伤的食品组合物,其中,所述食品为保健功能食品。
8.根据权利要求6所述的用于预防或改善肠道损伤的食品组合物,其中,所述食品为选自由面包、年糕、糖果、巧克力、口香糖、冰淇淋、牛奶、奶酪、加工肉制品、加工鱼制品、泡菜、酱油、大酱、辣椒酱、春酱、清国酱、醋、番茄酱、咖喱、调味汁、饮料及发酵油组成的组中的一种食品。
9.一种用于预防或改善肠道损伤的食品添加剂组合物,其中,包含柠檬明串珠菌菌株或其培养物作为有效成分。
10.一种用于预防或改善家畜的肠道损伤的饲料组合物,其中,包含柠檬明串珠菌菌株或其培养物作为有效成分。
11.根据权利要求10所述的用于预防或改善家畜的肠道损伤的饲料组合物,其中,所述家畜为选自由猪、牛、马、绵羊、兔、山羊、小鼠、仓鼠、豚鼠、狗、猫、鸡、火鸡、鸭、鹅、雉鸡、鹌鹑、鲤鱼、鲫鱼及鳟鱼而组成的组中的一种。
12.一种用于预防或改善家畜的肠道损伤的饲料添加剂组合物,其中,包含柠檬明串珠菌菌株或其培养物作为有效成分。
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KR10-2021-0017813 | 2021-02-08 | ||
KR10-2022-0014825 | 2022-02-04 | ||
KR1020220014825A KR20220114490A (ko) | 2021-02-08 | 2022-02-04 | 류코노스톡 시트래움 균주를 유효성분으로 포함하는 장 손상의 예방 또는 치료용 약학 조성물 |
PCT/KR2022/001870 WO2022169337A1 (ko) | 2021-02-08 | 2022-02-07 | 류코노스톡 시트래움 균주를 유효성분으로 포함하는 장 손상의 예방 또는 치료용 약학 조성물 |
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