WO2013177866A1 - 无指盘臭蛙促皮肤修复多肽ah90和cw49的应用 - Google Patents

无指盘臭蛙促皮肤修复多肽ah90和cw49的应用 Download PDF

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WO2013177866A1
WO2013177866A1 PCT/CN2012/080221 CN2012080221W WO2013177866A1 WO 2013177866 A1 WO2013177866 A1 WO 2013177866A1 CN 2012080221 W CN2012080221 W CN 2012080221W WO 2013177866 A1 WO2013177866 A1 WO 2013177866A1
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skin
repair
frog
slice
fingerless
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PCT/CN2012/080221
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French (fr)
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赖仞
杜彦军
容明强
刘涵
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中国科学院昆明动物研究所
四川科伦新光生物科技开发有限公司
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Publication of WO2013177866A1 publication Critical patent/WO2013177866A1/zh

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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
    • A61K35/56—Materials from animals other than mammals
    • A61K35/65—Amphibians, e.g. toads, frogs, salamanders or newts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like

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  • the invention belongs to the technical field of polypeptide medicine in biochemistry, and particularly relates to a skinless repairing polypeptide AH90 and CW49. Use in promoting skin repair, reducing scarring, and treating related skin diseases.
  • the skin acts as a natural protective barrier for the body and acts as a mechanical barrier to pathogens. Once the natural barrier function of the skin is damaged, the body will automatically open the physiological switch to repair the damage of the skin.
  • the healing of human skin after injury there are two processes of tissue recruitment and scar healing mentioned above.
  • the epithelial cells grow from the normal skin edge to the heart, or from the epithelium of the sweat glands and hair follicles of the skin appendage, and finally complete the epidermal cover.
  • the response of dermal tissue to injury is the formation of granulation tissue, which is actually an inflammatory tissue. From a cytological point of view, granulation tissue includes: neonatal capillary endothelial cells, differentiated from local primitive mesoderm cells.
  • Fibroblasts and their compositions such as collagen fibers, mucopolysaccharides, and the like.
  • Scars are formed by the development of granulation tissue. After a certain period of growth, they stop and gradually absorb and resolve. The scar is still proliferating within half a year to one year after the general wound healing, and then gradually absorbed. However, this process can not be completed for several years or more than ten years.
  • the fingerless stink frog is an amphibious animal of the frog family, and is a unique species in China. Distributed in Sichuan, Guizhou, Yunnan and other places, often living in the small hills and rivers in the lush vegetation and dark and humid places. Its survival range is 720 to 3200 Meter.
  • the fingerless stinky frog adapts to its natural environment and avoids injury such as injury.
  • the skin of the fingerless frog has developed into a multi-functional organ, such as: secreting a large number of antibacterial peptides against microorganisms. Invasion; secrete neurotoxin against various natural enemies; secrete growth factors to promote wound healing.
  • the non-finger odor frog of the present invention promotes the skin repair function of the skin repair polypeptides AH90 and CW49, and there has not been any public report that the two polypeptides can repair the skin.
  • the object of the present invention is to provide a skinless repair polypeptide AH90 and CW49
  • the invention relates to the preparation of a medicament for treating body surface wounds, burns, skin ulcers, reducing scar production and accelerating scar repair.
  • the fingerless frog-promoting skin repairing polypeptides AH90 and CW49 used in the present invention are obtained by a conventional chemical synthesis method; wherein:
  • the molecular weight of the skin repair peptide AH90 is 2651.18 Da, the isoelectric point is 10.26, and its full sequence primary structure is ATAWDFGPHGLLPIRPIRIRPLCG.
  • non-finger odor-prone skin repairing polypeptides AH90 and CW49 of the present invention for the preparation of a medicament for treating body surface wounds, burns, skin ulcers, reducing scar production, and accelerating scar repair.
  • the content of the present invention is tested on an animal model of mice, and the test protocol is a conventional method, as follows:
  • mice After the mice were anesthetized with 2% sodium pentobarbital, the skin on both sides of the back was depilated with a shaver. After alcohol disinfection, two wounds of 7 mm in diameter were opened behind the sample, and the sample was dissolved in PBS (concentration of AH 0.05 Mg/ml, CW concentration of 1 mg/ml), administered twice daily for 30 ul each time. The sample was applied to the right side of the wound, and the left side of the wound was filled with the corresponding volume of PBS as a negative control. The wound changes were taken with the camera every two days, namely DAY0, DAY2, DAY4, DAY6, DAY8 until the wounded wound was completely dislocated. When taking a photo, the measuring ruler is used as a ruler. Finally used as a graph software to calculate the change in wound size, using histological Score rating system score.
  • PBS concentration of AH 0.05 Mg/ml, CW concentration of 1 mg/ml
  • the fingerless stinky frog promotes the proliferative effect of the skin repair peptides AH90 and CW49 on the dermis and epidermis.
  • the wound skin tissue was cut out after the mice were sacrificed.
  • the cut liver tissue was washed with physiological saline tissue and immediately placed in a neutral formalin fixative.
  • the material is dehydrated by 70%, 80%, 90% ethanol solution, 30min each, then 95%, 100% 2 times each, 20 minutes each time.
  • Adjust the thickness adjuster to the desired slice thickness typically 4-10 microns.
  • the Lord can start slicing. At this time, shake the runner in the right hand, cut the wax block into a wax belt, and lift the wax belt with the brush on the left.
  • the rocking speed should not be too fast, usually 40-50r/min.
  • the slicing knife should be removed and the paraffin wax on the knife should be wiped off with chloroform, and the microtome should be wiped clean and stored.
  • the temperature of the water bath was adjusted to 60 ° C.
  • the slice and the slice holder were placed in a dry dyeing tank, placed in a water bath, and covered with a lid (sealable), and the wax was melted for 30 minutes.
  • the paraffin sections were dewaxed by xylene I and II for 5 min, then placed in 100%, 95%, 90%, 80%, 70% alcoholic solutions for 3-5 min, and then placed in distilled water for 3 min.
  • the sections were immersed in 1% hydrochloric acid in ethanol for 2 seconds to tens of seconds, and the slices turned red and the color was light.
  • the stained slices were photographed, and finally the thickness of the epidermis was calculated using the software ImageJ, using histological Score rating system score.
  • the fingerless scented frog promoting skin repairing polypeptide AH90 and CW49 of the present invention can promote the repair of the skin and reduce the hyperplasia of the epidermis during skin repair. It can be used to prepare drugs for treating surface wounds, burns and skin ulcers.
  • Figure 1-A, Figure 1-B, Figure 1-C, Figure 1-D, and Figure 1-E show 0 days, 2 days, 4 Days, 6 days, 8 days The control and therapeutic effect of AH90 on skin damage model in mice.
  • Figure 2-A, Figure 2-B, Figure 2-C, Figure 2-D, and Figure 2-E show 0 days, 2 days, 4 Days, 6 days, 8 days without the control and therapeutic effect of the skin-repairing polypeptide CW49 on the skin damage model of mice.
  • Figure 3-A, Figure 3-B, Figure 3-C, Figure 3-D, Figure 3-E, and Figure 3-F are respectively Fingerless odor frog to promote skin repair peptide AH90 on the mouse skin control and therapeutic effect slice.
  • Figure 4-A, Figure 4-B, Figure 4-C, Figure 4-D, Figure 4-E, and Figure 4-F are respectively A cross-sectional view of the control and therapeutic effect of CW49 on the skin of mice without the finger odor frog.
  • Figure 5-A and Figure 5-B show the skin repair peptides AH90 and CW49, respectively. A score for skin healing.
  • Figure 6-A and Figure 6-B show the skin repair peptides AH90 and CW49, respectively. Proliferation of the epidermis.
  • Figure 7-A and Figure 7-B show the skin repair peptides AH90 and CW49, respectively. Score the epidermal proliferation.
  • the fingerless scented frog-promoting polypeptide AH90 and CW49 used in the present invention Obtained by conventional chemical synthesis; among which:
  • No finger odor frog promotes skin repair peptide AH90 has a molecular weight of 2657.18 Da
  • the isoelectric point is 10.26, and the full sequence primary structure is ATAWDFGPHGLLPIRPIRIRPLCG.
  • the molecular weight of CW49 is 1210.43 Da and the isoelectric point is 8.29.
  • the full sequence primary structure is APFRMGICTTN.
  • the invention has no finger odor frog skin promoting polypeptide AH9 0 and CW49 in preparation Application in the treatment of body surface wounds, burns, and skin ulcers.
  • the content of the present invention is tested on an animal model of a mouse, and the test thereof
  • the scheme is a conventional method, and the specific steps are the same as those described in the section of the invention.
  • Fingerless odor frog promotes skin repair peptides AH90 and CW49 scores for granulation tissue healing.
  • Fingerless odor frog promotes skin repair polypeptide AH90 and CW49 on epidermal proliferation.
  • the thickness of the epidermis increased significantly during the recovery of mouse skin damage, smearing
  • the thickness of the epidermis after AH90 and CW49 was significantly lower than that of the control group and there was a statistically significant difference (*P ⁇ 0.01), AH90 and CW49 has the effect of preventing scarring.
  • Fingerless scented frogs promote skin repair peptides AH90 and CW49 score on dermal and epidermal proliferation.
  • the skin recovered from the mouse was sectioned, and then photographed to observe the proliferation thickness of the epidermis, using histological score
  • the scoring system scores skin repairs.
  • the results showed (Fig. 7-A, Fig. 7-B), using the finger odor frog to promote skin repair peptides AH90 and CW49 There was a significant difference between the epidermal and dermal scores and the control group.

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Abstract

提供了无指盘臭蛙促皮肤修复多肽AH90和CW49中的任意一种在制备治疗体表创伤、烧伤、皮肤溃疡、减少疤痕产生、加快疤痕修复药物中的应用。其中，多肽AH90的分子量为2657.18Da，等电点为10.26，全序列一级结构为ATAWDFGPHGLLPIRPIRIRPLCG；多肽CW49的分子量为1210.43Da，等电点为8.29，全序列一级结构为APFRMGICTTN。

Description

无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 的应用 技术领域
本发明属于生物化学中的多肽药物技术领域，具体涉及无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 在促进皮肤修复、减少疤痕以及治疗相关皮肤性疾病中的应用。
背景技术
皮肤作为机体天然的保护屏障，有机械阻挡病原体的作用。一旦皮肤的天然屏障作用被损坏，机体就会立即自动开启生理开关进行皮肤的损伤修复。人体皮肤损伤后的愈合，同时存在上述的组织补充和疤痕愈合两个过程。上皮细胞由正常皮肤边缘向心性爬行生长，或由皮肤附属器汗腺、毛囊的上皮向外扩展生长，最后完成创面上皮覆盖。真皮组织对损伤的反应，是形成肉芽组织，肉芽组织实际上是一种炎症组织，从细胞学角度来看，肉芽组织包括：新生的毛细血管内皮细胞，由局部原始的中胚层细胞分化来的纤维母细胞及其合成物，如胶原纤维、粘多糖等。疤痕是由肉芽组织发育形成，生长到一定阶段后自行停止并逐渐吸收消退。一般的创面愈合后半年至一年内瘢痕仍在增生，其后逐渐吸收平复。但此过程可延续几年、十几年尚不能完成。
无指盘臭蛙为蛙科蛙属的两栖动物，是中国的特有物种。分布于四川、贵州、云南等地，常生活于中小山溪内植物茂盛以及阴暗潮湿处。其生存的海拔范围为 720 至 3200 米。无指盘臭蛙适应其自生存的自然环境以及避免受伤等机体损伤，在长期的进化过程中，无指盘臭蛙的皮肤发展成为具有多种功能的器官，如：分泌大量抗菌肽抵御微生物侵袭；分泌神经毒素对抗各类天敌；分泌生长因子促进伤口愈合等。
本发明研究的无指盘臭蛙促皮肤修复多肽AH90和CW49对皮肤修复功能，目前为止尚未有这两种多肽能修复皮肤的公开报道。
 技术问题
本发明的目的在于提供无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 在制备治疗体表创伤、烧伤、皮肤溃疡、减少疤痕产生、加快疤痕修复药物中的应用。
技术解决方案
本发明的所用的无指盘臭蛙促皮肤修复多肽AH90和CW49采用常规的化学合成的方法获得；其中：
1. 无指盘臭蛙促皮肤修复多肽AH90的分子量为2657.18 Da，等电点为10.26，其全序列一级结构为ATAWDFGPHGLLPIRPIRIRPLCG。
2、无指盘臭蛙促皮肤修复多肽CW49的分子量为1210.43 Da，等电点为8.29，全序列一级结构为APFRMGICTTN。
本发明的无指盘臭蛙促皮肤修复多肽AH90和CW49在制备治疗体表创伤、烧伤、皮肤溃疡、减少疤痕产生、加快疤痕修复药物中的应用。
本发明的内容在小鼠的动物模型上试验，其试验方案为常规方法，具体如下：
I.无指盘臭蛙促皮肤修复多肽AH90和CW49对伤口的愈合作用
小鼠用2%戊巴比妥钠麻醉后,背部两侧皮肤用剃毛器除毛,酒精消毒后,在其后背开两个直径7mm的创口，样品溶解于PBS中（AH的浓度0.05mg/ml，CW浓度为1mg/ml）,每天给药两次，每次30ul。样品加在右侧创面，左侧创面加相应体积的PBS作阴性对照。每隔两天用相机拍下创面变化，即DAY0，DAY2，DAY4，DAY6，DAY8直到加药创面完全脱痂愈合为止。拍照时用量尺比照创口作标尺。最后用作图软件计算创面大小变化，采用histological score的评分系统评分。
II．无指盘臭蛙促皮肤修复多肽AH90和CW49对真皮、表皮的增生作用。
A:石蜡切片
1．取材
将小鼠处死后剪取创面皮肤组织。
2．固定
将切好的肝组织用生理盐水组织洗一下，立即投入中性福尔马林固定液中。
3. 洗涤
材料经固定后,流水冲洗，数小时或过夜。
4. 脱水
材料依次经70%、80%、90%各级乙醇溶液脱水，各30min，再放入95%、100% 各2次，每次20min。
5．透明
纯酒精、二甲苯等量混合液15min，二甲苯Ⅰ15min、Ⅱ15min（至透明为止）。
6．透蜡
放入二甲苯和石蜡各半的混合液15min，再放入石蜡Ⅰ、石蜡Ⅱ 透蜡各50-60分钟。
7. 包埋
用镊子夹取石蜡模子（金属质地）在酒精灯上稍加热，放在平的 桌面上，从温箱中取出盛放纯石蜡的蜡杯，倒入少许石蜡。再将镊子在酒精灯上 稍加热，夹取材料将切面朝下放入蜡模中，排列整齐。再放上包埋盒，轻轻倒入 熔蜡。
8. 切片
①将已固定和修好的石蜡块装在切片机的夹物台上。
②将切片刀固定在刀夹上，刀口向上。
③摇动推动螺旋，使石蜡块与刀口贴近，但不可超过刀口。
④调整石蜡块与刀口之间的角度与位置，刀片与石蜡切片约成15度左右。
⑤调整厚度调节器到所需的切片厚度，一般为4-10微米。
⑥一切调整好后主可以开始切片。此时右手摇动转轮，让蜡块切成蜡带，左手持毛笔将蜡带提起，摇转速度不可太急，通常以40-50r/min。
⑦切成的蜡带到20-30cm长时，右手用另一支毛笔轻轻将蜡带挑起，以免卷曲，并牵引成带，平放在蜡带盒上，靠刀面的一面较光滑，朝下，较皱的一面朝上。
⑧用单面刀片切取蜡片一小段，放在载玻上加水一滴，置于放大镜或显微镜下观察切片是否良好。
⑨切片工作结束后，应将切片刀取下用氯仿擦去刀上沾着的石蜡，把切片机擦拭干净保存。
9. 展片、贴片
打开水浴锅，使水温维持在40-45℃，另准备30%乙醇溶液。
① 切片时，将一碗30%乙醇溶液放于切片机旁的桌面上。
② 用小镊子夹取预先用刀片割开的蜡带，放在乙醇溶液的水面上，使切片展开。
③小镊子轻轻地将连在一起的切片分开，用一个载玻片将切片完整，已展开的切片捞至温水中，使之充分展开。
④ 另取洁净的载玻片，捞起展开的切片，使其位于切片1/3处，另一端（磨边，粗糙的一端）磨面上标记或贴上标签，放于切片架上。
10. 脱蜡复水
将水浴锅温度调至60℃，待水温控制在60℃时，将切片连同切片架放入一干燥的染色缸内，放入水浴锅中，盖上盖子（可密封），30min至蜡熔化。之后，石蜡切片经二甲苯Ⅰ、Ⅱ脱蜡各5min，然后放入100%、95%、90%、80%、70%各级酒精溶液中各3-5min，再放入蒸馏水中3min。
B:HE 染色
1.切片放入苏木精中染色约10-30min。 染色时间应根据染色剂的成熟程度及室温高低，适当缩短或延长。冬季室温低时放入恒温箱中染色。
2.水洗
用自来水流水冲洗15min。使切片颜色变蓝（或放入碱性水中），但要注意流水不能过大，以防切片脱落。
3．分化
将切片放入1%盐酸乙醇液中褪色，2秒至数十秒钟，见切片变红，颜色较浅。
4．漂洗
切片再放入自来水流水中使其恢复蓝色。
5．脱水Ⅰ
切片入50%乙醇→70%乙醇→80%乙醇中各3-5min。
6、复染 用0.5%伊红乙醇液对比染色1-3min。
7．脱水Ⅱ 将切片放入95%乙醇中洗去多余的红色，然后放入无水乙醇中3-5min。最后用吸水纸吸干多余的乙醇。
8．透明
切片放入二甲苯Ⅰ、Ⅱ中各3-5min。
9．封藏：中性树胶封存
C:表皮厚度计算。
将染色后的切片拍照，最后用软件ImageJ计算表皮厚度，采用histological score的评分系统评分。
 有益效果
本发明的 无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 能促进皮肤的修复，同时能减少皮肤修复过程中表皮的增生厚度， 能够用于 制备 治疗体表创伤、烧伤、皮肤溃疡的药物。
附图说明
图 1-A 、图 1-B 、图 1-C 、图 1-D 、图 1-E 分别显示 0 天、 2 天、 4 天、 6 天、 8 天 无指盘臭蛙促皮肤修复多肽 AH90 对小鼠皮肤损伤模型的对照及治疗作用。
图 2-A 、图 2-B 、图 2-C 、图 2-D 、图 2-E 分别显示 0 天、 2 天、 4 天、 6 天、 8 天无 指盘臭蛙促皮肤修复多肽 CW49 对小鼠皮肤损伤模型的对照及治疗作用。
图 3-A 、图 3-B 、图 3-C 、图 3-D 、图 3-E 、图 3-F 分别为 无指盘臭蛙促皮肤修复多肽 AH90 对小鼠皮肤的对照及治疗作用切片图。
图 4-A 、图 4-B 、图 4-C 、图 4-D 、图 4-E 、图 4-F 分别为 无指盘臭蛙促皮肤修复多肽 CW49 对小鼠皮肤的对照及治疗作用作用切片图。
图 5-A 、图 5-B 分别为 无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 对皮肤愈合的评分。
图 6-A 、图 6-B 分别为 无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 对表皮增殖作用。
图 7-A 、图 7-B 分别为无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 对表皮增殖评分。
 本发明的实施方式
本发明的所用的无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 采用常规的化学合成的方法获得；其中：
1. 无指盘臭蛙促皮肤修复多肽 AH90 的分子量为 2657.18 Da ，等电点为10.26，其全序列一级结构为 ATAWDFGPHGLLPIRPIRIRPLCG 。
2 、无指盘臭蛙促皮肤修复多肽 CW49 的分子量为 1210.43 Da ，等电点为 8.29 ， 全序列一级结构为 APFRMGICTTN 。
本发明的无指盘臭蛙促皮肤修复多肽AH9 0 和 CW49 在制备 治疗体表创伤、烧伤、皮肤溃疡药物中的应用。
本发明的内容在小鼠的动物模型上 试验 ，其 试验 方案为常规方法，具体步骤同发明内容部分的描述。
实施例 1 ：
无指盘臭蛙促皮肤修复多肽 AH90 对小鼠皮肤损伤模型的修复作用。
取 0.05mg/ml 无指盘臭蛙促皮肤修复多肽 AH90 进行试验，结果为：能迅速促进皮肤的修复功能（图 1-A 、图 1-B 、图 1-C 、图 1-D 、图 1-E ），第二天的修复效果比较明显，第四天结痂，第六天基本上完全恢复，并且和对照小鼠的皮肤修复有明显的 差异。
实施例 2 ：
无指盘臭蛙促皮肤修复多肽 CW49 对小鼠皮肤损伤模型的修复作用。
取 1mg/ml 无指盘臭蛙促皮肤修复多肽 CW49 进行试验，结果为：能迅速促进皮肤的修复功能（图 1-A 、图 1-B 、图 1-C 、图 1-D 、图 1-E ），第二天的修复效果比较明显，第四天结痂，第八天基本上完全恢复，并且和对照小鼠的皮肤修复有明显的 差异。
实施例 3 ：
无指盘臭蛙促皮肤修复多肽 AH90 对小鼠皮肤修复作用切片图。
小鼠皮肤在恢复的过程中，将每天恢复效果做成切片，观察皮肤表皮生长情况。 图 3-A 、图 3-B 、图 3-C 、图 3-D 、图 3-E 、图 3-F 显示加入 AH90 治疗后的第三天肉芽组织形成，第五天表皮组织基本上完全恢复，第九天治疗组的表皮组织厚度与对照组相比明显减少。
实施例 4 ：
无指盘臭蛙促皮肤修复多肽 CW49 对小鼠皮肤修复作用切片图。
切片结果表明（ 图 4-A 、图 4-B 、图 4-C 、图 4-D 、图 4-E 、图 4-F ），无指盘臭蛙促皮肤修复多肽 CW49 治疗后第三天形成较厚的增生表皮组织，第五天增生的表皮组织厚度减少，第九天表皮厚度恢复正常且有腺体产生。
实施例 5 ：
无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 对肉芽组织愈合的评分。
小鼠皮肤的创伤后涂抹 无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 ，对每天小鼠伤口的愈合情况拍照，采用 histological score 的评分系统对肉芽组织愈合进行评分。结果表明（图 5-A 、图 5-B ），使用 无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 后，肉芽组织愈合的程度和对照有明显 统计学差异（ *P<0.01 ）。
实施例 6 ：
无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 对表皮增殖作用。
如图 6-A 、图 6-B 所示，小鼠皮肤损伤恢复的过程中表皮的厚度明显增加， 涂抹 无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 后表皮的厚度增加明显小于对照组并且和对照组有明显 统计学差异（ *P<0.01 ） ， AH90 和 CW49 具有预防疤痕产生的作用。
实施例 7 ：
无指盘臭蛙促皮肤修复多肽 AH90 和 CW49 对真皮和表皮增殖评分。
将小鼠皮肤恢复的部位做成切片，然后拍照观察表皮的增殖厚度，采用 histological score 的评分系统对皮肤修复进行评分。结果表明（图 7-A 、图 7-B ），使用指盘臭蛙促皮肤修复多肽 AH90 和 CW49 表皮和真皮评分和对照组有显著性的差异。

Claims (2)

  1. 分子量为 2657.18 Da ，等电点为10.26，其全序列一级结构为 ATAWDFGPHGLLPIRPIRIRPLCG 的无指盘臭蛙促皮肤修复多肽 AH90 在制备治疗体表创伤、烧伤、皮肤溃疡、减少疤痕产生、加快疤痕修复药物中的应用。
  2. 分子量为 1210.43 Da ，等电点为 8.29 ， 全序列一级结构为 APFRMGICTTN 的无指盘臭蛙促皮肤修复多肽 CW49 在制备治疗体表创伤、烧伤、皮肤溃疡、减少疤痕产生、加快疤痕修复药物中的应用。
PCT/CN2012/080221 2012-05-31 2012-08-16 无指盘臭蛙促皮肤修复多肽ah90和cw49的应用 WO2013177866A1 (zh)

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