WO2019095685A1 - 一种功能多肽及其在制备防治肺纤维化药物中的应用 - Google Patents

一种功能多肽及其在制备防治肺纤维化药物中的应用 Download PDF

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WO2019095685A1
WO2019095685A1 PCT/CN2018/091731 CN2018091731W WO2019095685A1 WO 2019095685 A1 WO2019095685 A1 WO 2019095685A1 CN 2018091731 W CN2018091731 W CN 2018091731W WO 2019095685 A1 WO2019095685 A1 WO 2019095685A1
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pulmonary fibrosis
functional polypeptide
treatment
polypeptide
functional
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PCT/CN2018/091731
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French (fr)
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张海涛
余华军
伍俊
罗辉
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广东医科大学
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Priority to EP18865340.6A priority Critical patent/EP3553076A4/en
Priority to JP2019570612A priority patent/JP6826268B2/ja
Priority to US16/331,995 priority patent/US11225504B2/en
Priority to AU2018353935A priority patent/AU2018353935B2/en
Publication of WO2019095685A1 publication Critical patent/WO2019095685A1/zh

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4418—Non condensed pyridines; Hydrogenated derivatives thereof having a carbocyclic group directly attached to the heterocyclic ring, e.g. cyproheptadine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/10—Peptides having 12 to 20 amino acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the invention belongs to the technical field of medicine, and particularly relates to a functional polypeptide and application thereof in preparing a medicament for preventing and treating pulmonary fibrosis.
  • Idiopathic pulmonary fibrosis is an unexplained pulmonary interstitial inflammatory disease.
  • IPF Idiopathic pulmonary fibrosis
  • UPF common interstitial pneumonia
  • the main pathological features of IPF are mixed interstitial fibrosis and alveolar fibrosis and inflammatory cell infiltration, and progress to end-stage respiratory failure or death within 3 to 8 years after symptoms appear.
  • the etiology and pathogenesis of IPF have not yet been fully elucidated, lacking effective therapeutic drugs and objective and decisive prognostic indicators.
  • cytotoxic drugs are still the main treatment for IPF, but less than 30% of patients have a therapeutic response, but the drug shows toxic side effects.
  • the guidelines suggest that the vast majority of patients should not be treated with glucocorticoids, cyclosporine A, colchicine, IFN- ⁇ , bosentan and etanercept alone, nor do they recommend glucocorticoids and immunity. Inhibitor combination therapy.
  • pirfenidone for the treatment of IPF, which is currently used to treat mild to moderate IPF.
  • IPF idiopathic pulmonary fibrosis
  • side effects such as elevated transaminase, photosensitivity, eczema, abdominal discomfort, and decreased white blood cells.
  • the present invention aims to provide a functional polypeptide; it can significantly inhibit bleomycin-induced pulmonary fibrosis formation, and has a remarkable function of preventing and treating pulmonary fibrosis.
  • the present invention and the functional polypeptide are one or more of the 1, 2, 8, 9, 11, 13, and 14 positions based on the amino acid sequence shown in SEQ ID No. 1.
  • the amino group is substituted with a polypeptide having the same or similar functional amino group.
  • the invention also provides a coding nucleotide sequence identical to the functional polypeptide sequence.
  • the invention also provides a vector comprising the encoded nucleotide sequence.
  • the invention also provides a host cell comprising the vector.
  • the invention also provides the use of the functional polypeptide, which can be used for preparing a medicament for preventing and treating pulmonary fibrosis, or as an active ingredient for preventing and treating pulmonary fibrosis, or using the functional polypeptide as a precursor for preventing and treating pulmonary fibrosis, or Treatment of pulmonary fibrosis with a combination of pulmonary fibrosis drugs.
  • the specific plan is as follows:
  • a medicament for the treatment of pulmonary fibrosis comprising the polypeptide of the invention.
  • the functional polypeptide of the present invention has a good effect of preventing and treating IPF, and the content of hydroxyproline (HYP) and plasma malondialdehyde (MDA) in lung tissue is significantly decreased; total superoxide disproportionation in plasma The content of enzyme (T-SOD) glutathione peroxidase (GSH-PX) is significantly increased;
  • the functional polypeptide of the present invention can prepare an active ingredient or a prodrug of a drug or a drug for preventing and treating IPF;
  • the medicine for preventing and treating pulmonary fibrosis according to the present invention contains a functional polypeptide, which can be used as a powder or an injection or an oral product, and can be used as a finished product alone or as an active ingredient and physiological saline.
  • An adjuvant such as a glucose solution is formulated into a synthetic composition, and can also be used in combination with a currently used therapeutic pulmonary fibrosis drug.
  • Figure 1 is a LC-MS/MS analysis of the functional polypeptide structure of the present invention
  • Figure 3 is a graph showing the results of analysis of antibacterial activity of the functional polypeptide of the present invention.
  • Figure 4 is a graph showing the effect of the functional polypeptide of the present invention on the growth of human normal lung epithelial cells BEAS-2B;
  • Figure 5 is a graph showing the results of HE staining ( ⁇ 200) of mouse lung tissue sections in the IPF test for inhibition of EZY-1 polypeptide; wherein, 1. control group; 2. model group; 3.0.25 mg/kg EZY-1 polypeptide compound dose group; 4.0.5 mg/kg EZY-1 polypeptide compound dose group; 5.1.0 mg/kg EZY-1 polypeptide compound dose group; 6.50 mg/kg pirfenidone group.
  • the present invention provides a functional polypeptide which is
  • sequence of SEQ ID No. 1 is RTGACFCVIYNGILYP.
  • the present invention and the functional polypeptide are one or more of the 1, 2, 8, 9, 11, 13, and 14 positions based on the amino acid sequence shown in SEQ ID No. 1.
  • Amino groups are substituted with polypeptides having the same or similar functional amino groups. Specifically, it may be selected, but not limited to, one or more of the following substitutions: the first amino acid R in the sequence of SEQ ID No.
  • the second amino acid T is replaced by the amino acid S, because T and S are both hydroxyl-containing amino acids, which are functionally similar;
  • the eighth amino acid V is replaced by the amino group I or L, for the reason that V, I and L are both branched-chain amino acids, which are functionally similar;
  • the 9th amino acid I is replaced by V or L, on the grounds that V, I and L are both branched-chain amino acids, which are functionally similar;
  • the 11-position amino acid N is replaced by Q, because the N and Q are amino acid-containing amino acid groups, which are functionally similar;
  • the 13th amino acid I is replaced by V or L, for the reason that V, I and L are the same Branched-chain amino acids are functionally similar;
  • the 14th amino acid L is replaced by V or I, on the grounds that V, I and L are both branched-chain amino acids, which are functionally similar.
  • the invention also provides a coding nucleotide sequence identical to the functional polypeptide sequence.
  • the invention also provides a vector comprising the encoded nucleotide sequence.
  • the invention also provides a host cell comprising the vector.
  • the invention also provides the use of the functional polypeptide, which can be used for preparing a medicament for preventing and treating pulmonary fibrosis, or as an active ingredient for preventing and treating pulmonary fibrosis (IPF) drugs, or using the functional polypeptide as a medicament for preventing and treating pulmonary fibrosis (IPF) Precursor modification, or in combination with treatment of pulmonary fibrosis (IPF) drugs for the treatment of pulmonary fibrosis.
  • IPF pulmonary fibrosis
  • the specific plan is as follows:
  • IPF pulmonary fibrosis
  • the functional polypeptide compounds of the present invention can be used in the prevention and treatment of IPF in the following ways:
  • Method 1 The functional polypeptide compound of the present invention is orally administered at a dose of 1 to 10 mg/kg once a day for at least 35 days.
  • Method 2 The functional polypeptide compound of the present invention is injected at 0.1 to 1 mg/kg once every 3 days for continuous administration for at least 35 days.
  • Method 3 In combination with a currently used therapeutic IPF drug (for example, pirfenidone), the EZY-1 polypeptide compound is administered orally in an amount of 1 to 10 mg/kg once a day.
  • a currently used therapeutic IPF drug for example, pirfenidone
  • a medicament for the treatment of pulmonary fibrosis comprising the polypeptide of the invention.
  • the plurality of functions described in the present invention can be synthesized by amino acid as a raw material, synthesized by a peptide synthesis apparatus according to the amino acid sequence of SEQ ID No. 1, and purified by high performance liquid chromatography. The product was tested; the results are shown in Figures 1-4.
  • FIG. 1 is LC-MS/MS analysis of the functional polypeptide structure of the present invention
  • FIG. 2 is a result of SDS-PAGE electrophoresis analysis of the functional polypeptide of the present invention
  • FIG. 3 is an antibacterial activity of the functional polypeptide of the present invention. The result of the analysis shows that the functional polypeptide has a certain bacteriostatic effect
  • FIG. 4 is a graph showing the effect of the functional polypeptide of the present invention on the growth of human normal lung epithelial cells BEAS-2B, and the results show that the function of the present invention The polypeptide is less toxic to normal cells.
  • mice The random number table of mice was divided into the following groups: control group, model group, pirfenidone group (50 mg/kg), EZY-1 polypeptide low dose group (0.25 mg/kg), EZY-1 polypeptide medium dose group. (0.5 mg/kg), high dose group of EZY-1 polypeptide (1.0 mg/kg), 10 rats in each group.
  • mice were anesthetized on the back of the console.
  • the tongue was pulled out with tweezers to expose the trachea, and 5 mg/kg Bole was slowly injected under a cold light source magnifying glass. Mycin.
  • the rat plate was erected and rotated for 3 minutes to evenly distribute the drug solution in the lungs.
  • the control group was injected with an equal volume of isotonic saline in the same way as the previous method.
  • the rats were intragastrically administered with 0.1 ml/10 g.
  • the control group and the model group were intragastrically administered with an equal volume of isotonic saline.
  • the other groups were intragastrically administered with the corresponding dose of the drug for 35 days, once a day.
  • mice were anesthetized and sacrificed, and the intact lung tissues were isolated, weighed, and the same part of the right upper lobe was taken, fixed with 4% paraformaldehyde for 48 hours, and paraffin sections were taken for hematoxylin- Eosin staining (HE) staining for histopathological observation.
  • HE hematoxylin- Eosin staining
  • HYP Hydroxyproline
  • Table 1 show the plasma T-SOD, MDA, GSH-PX content of each group of mice.
  • the plasma MDA content in the model, low-medium and high-dose groups increased (P ⁇ 0.01).
  • the plasma T-SOD and GSH-PX levels in each group decreased (P ⁇ 0.01).
  • the plasma MDA content of each group was significantly decreased (P ⁇ 0.01).
  • the plasma T-SOD and GSH-PX levels of each group increased (P ⁇ 0.01).
  • the lung structure of the control group was clear, the alveolar space of the model group was significantly widened, the alveolar structure was severely damaged, the atrophy collapsed severely, the collagen fibers increased significantly, and a large number of inflammatory cells infiltrated.
  • the low, medium and high dose groups of EZY-1 peptide and pirfenidone group can reduce alveolar structure damage, reduce collagen fiber production, and inhibit inflammatory cell infiltration. It can be seen that the drug group gradually reduced pulmonary fibrosis.

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Abstract

本发明公开了一种功能多肽，其是1)具有SEQ ID No.1所述的氨基酸序列的多肽；或者2)具有在所述SEQ ID No.1所示的氨基酸序列基础上经过取代、缺失、或者添加一个或几个氨基酸后且具有相同或相似功能的多肽。本发明还公开了所述功能多肽的用途，所述功能多肽具有防治肺纤维化的功能。

Description

一种功能多肽及其在制备防治肺纤维化药物中的应用 技术领域
本发明属于医药技术领域，具体涉及一种功能多肽及其在制备防治肺纤维化药物中的应用。
背景技术
特发性肺纤维化(idiopathic pulmonary fibrosis，IPF)是一种不明原因的肺间质炎症性疾病，2011年IPF诊治国际指南中，将IPF定义为:一种慢性、进行性和纤维化性的间质性肺炎，组织学和(或)胸部高分辨CT特征性表现为普通性间质性肺炎(UIP)。IPF主要病理特点为肺间质和肺泡腔内纤维化和炎细胞浸润混合存在，多在出现症状3～8年内进展至终末期呼吸衰竭或死亡。IPF病因及发病机制目前还没有完全阐明，缺乏有效的治疗药物和客观的、决定性的预后指标。皮质激素或免疫抑制剂、细胞毒药物仍是IPF主要的治疗药物，但不足30％的病人有治疗反应，但药物却表现毒副反应。对IPF的治疗，指南提出绝大多数患者不应当单用糖皮质激素、环孢素A、秋水仙碱、IFN-β、波生坦和依那西普治疗，也不推荐糖皮质激素与免疫抑制剂联合治疗。
欧盟、日本、加拿大，中国已批准使用吡非尼酮治疗IPF，目前该药用于治疗轻中度IPF。尽管吡非尼酮已经比准上市，但目前对于吡非尼酮治疗特发性肺纤维化(IPF)还存在一定的争议，并且吡非尼酮治疗过程中会出现光敏反应、厌食、头晕、转氨酶升高、光敏反应、湿疹、腹部不适和白细胞降低等副作用。2015年，美国胸科学会(ATS)/欧洲呼吸学会(ERS)/日本呼吸学会(JRS)/拉丁美洲胸科学会(ALAT)联合在IPF治疗临床实践指南建议使用吡非尼酮对IPF患者进 行治疗(有条件推荐，中等级别证据)。理由和实施注意事项：吡啡尼酮的不良反应较广，即便是FVC评估治疗受益的情况下，部分患者仍不能忍受某种不良反应，因此开始治疗前，应就不良反应进行患者教育。此外，目前吡啡尼酮治疗费用较高，在做出治疗决定时，特别是患者直接承担治疗的经济负担时，必须将这一影响因素考虑进去。无论在欧洲还是日本使用吡非尼酮价格都较高，虽然此药已在临床上使用，但其改善IPF患者的症状或生活质量，以及降低患者死亡率的效果有限。在今后仍需要通过已上市国家和地区的临床观察结果给出确切的结论。因此寻找新的防治IPF有效药物仍然是国内外药物研究的热点。
发明内容
为了克服现有技术的不足，本发明的目的在于提供一种功能多肽；能够显著地抑制博来霉素诱导的肺纤维化形成，具有显著的防治肺纤维化功能。
为解决上述问题，本发明所采用的技术方案如下：
一种功能多肽，其是
1)具有SEQ ID No.1所述的氨基酸序列的多肽；或者
2)具有在所述SEQ ID No.1所示的氨基酸序列基础上经过取代、缺失、或者添加一个或几个氨基酸后且具有相同或相似功能的多肽。
作为进一步的方案，本发明和所述的功能多肽是在所述SEQ ID No.1所示的氨基酸序列基础上第1、2、8、9、11、13、14位中的一个或多个氨基被取代成具有相同或相似功能氨基的多肽。
本发明还提供了一种与所述功能多肽序列相同的编码核苷酸序列。
本发明还提供了一种含有所述的编码核苷酸序列的载体。
本发明还提供了一种含有所述的载体的宿主细胞。
本发明还提供了所述功能多肽的用途，其可用于制备防治肺纤维化药物，或作为防治肺纤维化药物的有效成分，或利用该功能多肽作为防治肺纤维化药物前体改造，或与治疗肺纤维化药物联合应用治疗肺纤维化。具体方案如下：
一种如本发明所述的功能多肽在制备防治肺纤维化药物中的应用。
一种如本发明所述的功能多肽在制备防治肺纤维化药物的有效成分中的应用。
一种如本发明所述的功能多肽在制备防治肺纤维化药物的前体重的应用。
一种如本发明所述的功能多肽与吡非尼酮联用在制备防治肺纤维化药物中的应用。
一种治疗防治肺纤维化药物，包含本发明所述的多肽。
相比现有技术，本发明的有益效果在于：
1.本发明所述的功能多肽的具有很好的防治IPF的效果，肺组织中羟脯氨酸(HYP)及血浆中丙二醛(MDA)的含量明显降低；血浆中总超氧化物歧化酶(T-SOD)谷胱甘肽过氧化物酶(GSH-PX)的含量明显增加；
2.本发明所述的功能多肽可以制备防治IPF的药物或药物的有效成分或药物前体；
3.本发明所述的防治肺纤维化的药物、保健品含有功能多肽，能够制成粉剂或注射针剂或口服产品等多种形式，可以单独作为成品使用，也可以作为有效成分与生理盐水、葡萄糖溶液等佐剂配合成组合物，还可以与目前使用的治疗肺纤维化药物联合使用。
下面结合附图及具体实施方式对本发明作进一步详细说明。
附图说明
图1为LC-MS/MS分析本发明所述的功能多肽结构；
图2为SDS-PAGE电泳分析本发明所述的功能多肽结果图；
图3为本发明所述的功能多肽抗菌活性分析结果图；
图4为本发明所述的功能多肽对人正常肺上皮细胞BEAS-2B生长影响结果分析图；
图5为EZY-1多肽抑制IPF试验中小鼠肺组织切片HE染色观察(×200)结果图；其中，1.对照组；2.模型组；3.0.25mg/kg EZY-1多肽化合物剂量组；4.0.5mg/kg EZY-1多肽化合物剂量组；5.1.0mg/kg EZY-1多肽化合物剂量组；6.50mg/kg吡非尼酮组。
具体实施方式
本发明提供一种功能多肽，其是
1)具有SEQ ID No.1所述的氨基酸序列的多肽；或者
2)具有在所述SEQ ID No.1所示的氨基酸序列基础上经过取代、缺失、或者添加一个或几个氨基酸后且具有相同或相似功能的多肽；
其中SEQ ID No.1序列为RTGACFCVIYNGILYP。
作为进一步的方案，本发明和所述的功能多肽是在所述SEQ ID No.1所示的氨基酸序列基础上第1、2、8、9、11、13、14位中的一种或多种氨基经被取代成具有相同或相似功能氨基的多肽。具体可以选择但不限于下述一种方式或多种方式的取代：将SEQ ID No.1序列中第1位氨基酸R替换成K氨基，理由在于R和K同为碱性氨基酸，在功能上具有相似性；将第2位氨基酸T换为氨基S，理由在于T和S同为含羟基氨基酸，在功能上具有相似性；将第8位氨基酸V替换为氨基I或L，理由是V、I和L同为支链氨基酸，在功能上具有相 似性；将第9位氨基酸I替换为V或L，理由是V、I和L同为支链氨基酸，在功能上具有相似性；将第11位氨基酸N替换为Q，理由是N和Q同为含氨酰胺基团氨基酸，在功能上具有相似性；件第13位氨基酸I替换为V或L，理由是V、I和L同为支链氨基酸，在功能上具有相似性；将第14位氨基酸L替换为V或I，理由是V、I和L同为支链氨基酸，在功能上具有相似性。
本发明还提供了一种与所述功能多肽序列相同的编码核苷酸序列。
本发明还提供了一种含有所述的编码核苷酸序列的载体。
本发明还提供了一种含有所述的载体的宿主细胞。
本发明还提供了所述功能多肽的用途，其可用于制备防治肺纤维化药物，或作为防治肺纤维化(IPF)药物的有效成分，或利用该功能多肽作为防治肺纤维化(IPF)药物前体改造，或与治疗肺纤维化(IPF)药物联合应用治疗肺纤维化。具体方案如下：
一种如本发明所述的功能多肽在制备防治肺纤维化药物(IPF)中的应用。
一种如本发明所述的功能多肽在制备防治肺纤维化(IPF)药物的有效成分中的应用。
一种如本发明所述的功能多肽在制备防治肺纤维化(IPF)药物的前体重的应用。
一种如本发明所述的功能多肽与吡非尼酮联用在制备防治肺纤维化(IPF)药物中的应用。
本发明所述的功能多肽化合物在防治IPF的应用可以有如下多种方法：
方法1：口服本发明的功能多肽化合物1～10mg/kg，每天一次，连续服药至少35天。
方法2：注射本发明的功能多肽化合物0.1～1mg/kg，每3天一次，连续服 药至少35天。
方法3：与目前使用的治疗IPF药物(例如吡非尼酮)联合应用，口服EZY-1多肽化合物1～10mg/kg，每天一次。
一种治疗防治肺纤维化药物，包含本发明所述的多肽。
以下是本发明具体的实施例，在下述实施例中所采用的原料、试剂等除在本发明中有特殊限定的，均为现有技术。
本发明所述的功能多条可以以氨基酸为原料，用多肽合成仪器，按照SEQ ID No.1所述的氨基酸序列人工合成，用高效液相色谱纯化。并对产物进行检测；检测结果参见图1-4。
其中，图1为LC-MS/MS分析本发明所述的功能多肽结构；图2为SDS-PAGE电泳分析本发明所述的功能多肽结果图；图3为本发明所述的功能多肽抗菌活性分析结果图，结果显示该功能多肽具有一定的抑菌作用；图4为本发明所述的功能多肽对人正常肺上皮细胞BEAS-2B生长影响结果分析图，结果显示，本发明所述的功能多肽对正常细胞毒性低。
防治肺纤维化IPF实验
1、试验方法
(1)动物：SPF级C57小鼠，22g±2g，60只，雌性。
(2)分组
小鼠随机数字表法分为以下各组：对照组、模型组、吡非尼酮组(50mg/kg)、EZY-1多肽低剂量组(0.25mg/kg)、EZY-1多肽中剂量组(0.5mg/kg)、EZY-1多肽高剂量组(1.0mg/kg)，每组10只。
(3)用1％戊巴比妥钠按照50mg/kg的剂量腹腔注射麻醉小鼠后，仰卧固定于操作台，用镊子拉出舌头暴露气管，在冷光源放大镜下缓慢注入5mg/kg博 莱霉素。注射完毕后迅速竖起鼠板，旋转3分钟，使药液在肺内均匀分布。对照组同前方法气管内注入等体积等渗盐水。10天后开始以0.1ml/10g进行灌胃给药，对照组及模型组灌胃等体积的等渗盐水，其余各组灌胃相应剂量的药物，连续给药至35天，每天1次。
(4)取材与指标测定：
给药35天后将小鼠麻醉取血处死，分离得到完整的两肺组织，称重，取右肺上叶同一部位，用4％的多聚甲醛固定48小时，石蜡切片，进行苏木精-伊红染色法(HE)染色供组织病理学观察。
取左肺中叶同一部位进行肺组织HYP含量的检测。
收集血浆，检测T-SOD、MDA、GSH-PX含量。
统计学分析应用SPSS 17.0软件进行统计分析，计量资料用均数±标准差(x±s)表示，多个样本均数的比较采用单因素方差分析，组间比较采用t检验，以P≤0.05表示为差异有统计学意义。
2、结果
(1)小鼠肺组织HYP含量分析
各组小鼠肺组织羟脯氨酸(HYP)含量
Figure PCTCN2018091731-appb-000001
与对照组相比，各组小鼠肺组织HYP含量上升(P＜0.01)；与模型组相比，各组HYP含量明显下降(P＜0.01)。见表1。
(2)小鼠血浆T-SOD、MDA、GSH-PX含量分析，结果参见表1。
表1的结果显示：各组小鼠血浆T-SOD、MDA、GSH-PX含量
Figure PCTCN2018091731-appb-000002
与对照组相比，模型、低中、高剂量组小鼠血浆MDA含量上升(P＜0.01)；与对照组相比，各组小鼠血浆T-SOD、GSH-PX含量下降(P＜0.01)。与模型组相比，各组小鼠血浆MDA含量明显下降(P＜0.01)；与模型组相比，各组小鼠血浆 T-SOD、GSH-PX含量上升(P＜0.01)。
表1 各组生化指标比较
Figure PCTCN2018091731-appb-000003
与正常组比较：＊P＜0.01；与模型组比较：#p＜0.01，
表1的结果显示本发明所述的功能多肽能减轻胶原纤维沉着，抑制肺纤维化。
(3)肺组织病理变化分析，结果参见图5。
如图5，对照组肺结构清晰，模型组肺泡间隔显著变宽，肺泡结构破坏严重，萎缩塌陷严重，胶原纤维显著增多，出现大量炎症细胞浸润。而EZY-1多肽低、中、高剂量组及吡非尼酮组可以减轻肺泡结构破坏，减少胶原纤维生成，抑制炎症细胞浸润，可见药物组使肺纤维化逐步减轻。
上述实施方式仅为本发明的优选实施方式，不能以此来限定本发明保护的范围，本领域的技术人员在本发明的基础上所做的任何非实质性的变化及替换均属于本发明所要求保护的范围。

Claims (10)

  1. 一种功能多肽，其特征在于，其是
    1)具有SEQ ID No.1所述的氨基酸序列的多肽；或者
    2)具有在所述SEQ ID No.1所示的氨基酸序列基础上经过取代、缺失、或者添加一个或几个氨基酸后且具有相同或相似功能的多肽。
  2. 如权利要求1所述的功能多肽，其特征在于，其是在所述SEQ ID No.1所示的氨基酸序列基础上第1、2、8、9、11、13、14位中的一个或多个氨基被取代成具有相同或相似功能氨基的多肽。
  3. 一种与权利要求1所述的功能多肽序列相同的编码核苷酸序列。
  4. 含有权利要求3所述的编码核苷酸序列的载体。
  5. 含有权利要求4所述的载体的宿主细胞。
  6. 一种如权利要求1所述的功能多肽在制备防治肺纤维化药物中的应用。
  7. 一种如权利要求1所述的功能多肽在制备防治肺纤维化药物的有效成分中的应用。
  8. 一种如权利要求1所述的功能多肽在制备防治肺纤维化药物的前体中的应用。
  9. 一种如权利要求1所述的功能多肽与吡非尼酮联用在制备防治肺纤维化药物中的应用。
  10. 一种治疗防治肺纤维化药物，其特征在于，包含如权利要求1所述的多肽。
PCT/CN2018/091731 2017-11-17 2018-06-16 一种功能多肽及其在制备防治肺纤维化药物中的应用 WO2019095685A1 (zh)

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