CN111808050B - 混源萜penicimeroterpenoids A–C及其制备方法和应用 - Google Patents
混源萜penicimeroterpenoids A–C及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了混源萜penicimeroterpenoids A–C及其制备方法和应用。所述的混源萜类化合物Penicimeroterpenoids A、B、C的结构式如式(Ⅰ)所示。化合物Penicimeroterpenoids A、B、C是从Penicilliumsp.SCSIO 41509的发酵液和菌丝体中分离得到的，经试验证明，本发明的3个化合物Penicimeroterpenoids A、B、C具有抑制胰蛋白酶、人白细胞弹性蛋白酶、蛋白酪氨酸磷酸酶、蛋白酪氨酸磷酸酶SHP1的活性，它们可用于胰蛋白酶抑制剂、人白细胞弹性蛋白酶抑制剂、蛋白酪氨酸磷酸酶CDC25B抑制剂、蛋白酪氨酸磷酸酶SHP1抑制剂先导化合物的研究。
Description
技术领域
本发明属于海洋生物领域，具体涉及3个混源萜类化合物penicimeroterpenoidsA–C及其制备方法和在制备胰蛋白酶抑制剂、人白细胞弹性蛋白酶抑制剂、蛋白酪氨酸磷酸酶CDC25B抑制剂、蛋白酪氨酸磷酸酶SHP1抑制剂中的应用。
背景技术
胰蛋白酶的缺乏会导致许多人类疾病，如癌症、心血管疾病和炎症性疾病，胰蛋白酶抑制剂可用于抗癌、抗炎、治疗心血管疾病的药物研究中。人白细胞弹性蛋白酶是抗炎药物开发的主要靶点，人白细胞弹性蛋白酶抑制剂可用于抗炎药物研究中。而一些蛋白酪氨酸磷酸酶PTPs，如SHP1、CDC25B和TCPTP是潜在的抗癌药物靶点。SHP1和SHP2均与免疫疾病、癌症、神经疾病、感染性疾病有密切关系。SHP1和SHP2酶抑制剂可以用于制备治疗免疫疾病、癌症、神经疾病、感染性疾病的药物中应用。
海洋来源真菌能产生结构新颖、独特的次生代谢产物，是众多尚未被发掘和利用的新颖生物活性物质的重要生产源，是人类寻找和开发新型功能产物和天然药物的宝库。从海洋来源真菌中发掘小分子胰蛋白酶抑制剂、胰蛋白酶抑制剂和PTPs抑制剂，对于开发可用于治疗癌症和炎症等人类重大疾病方面的药物具有潜在重要应用价值。
发明内容
本发明的第一个目的是提供对胰蛋白酶、人白细胞弹性蛋白酶、蛋白酪氨酸磷酸酶CDC25B、蛋白酪氨酸磷酸酶SHP1有抑制作用的3个新混源萜类化合物penicimeroterpenoids A–C。
本发明3个新混源萜类化合物penicimeroterpenoids A–C，其结构式如式(Ⅰ)所示，
本发明的第二个目的是提供化合物Penicimeroterpenoids A、B、C的制备方法，它们是从真菌Penicilliumsp.SCSIO 41512的发酵产物中分离得到的。
优选，具体步骤如下：
(a)制备真菌Penicilliumsp.SCSIO 41512的发酵产物；
(b)将步骤(a)得到的发酵液用大孔树脂吸附，接着用水冲洗大孔树脂去掉培养基成分，再用甲醇或乙醇冲洗大孔树脂，浓缩有机相得到甲醇或乙醇提取物；或将步骤(a)得到的发酵液用乙酸乙酯、二氯甲烷或氯仿溶剂萃取，浓缩得到乙酸乙酯提取物、二氯甲烷提取物或氯仿提取物；菌丝体破碎后用丙酮浸泡，提取液减压浓缩除去丙酮，剩余水相用乙酸乙酯、二氯甲烷或氯仿萃取，浓缩得菌丝体的乙酸乙酯、二氯甲烷或氯仿提取物；合并菌丝体和发酵液的提取物，得到总提取物；
(c)将步骤(b)的总提取物经过正相硅胶柱层析，依次用二氯甲烷:甲醇体积比为100:0,95:5,90:10,80:20,70:30,60:40,50:50的溶剂系统梯度洗脱，收集二氯甲烷:甲醇体积比95:5冲洗下来的样品得到组分Fr.2；组分Fr.2通过中压反相ODS柱分离，以甲醇/水/TFA体积比5:95:0.03-100:0:0.03梯度洗脱，其中用甲醇/水/TFA(v/v/v,65:35:0.03)梯度洗脱得到的组分Fr.2-7经过凝胶Sephadex LH-20柱分离，以甲醇为洗脱溶剂，结合TLC，以二氯甲烷：甲醇＝v/v20:1为展开剂，分别收集Rf为0.3的流份为组分Fr.2-7-2，Rf为0.4的流份为Fr.2-7-1，Fr.2-7-1进一步通过半制备型HPLC纯化，以甲醇/水/TFA(v/v/v,70:30:0.03)等度洗脱得到化合物Penicimeroterpenoid A，Fr.2-7-2通过半制备型HPLC纯化，以甲醇/水/TFA(v/v/v,65:35:0.03)等度洗脱得到化合物PenicimeroterpenoidsB和C。
进一步优选，步骤(a)中所述的发酵产物是通过以下方法制备：将真菌Penicilliumsp.SCSIO 41512在真菌适用的平板培养基中生长，待真菌长出孢子后，将真菌接种到发酵培养基中，于室温静置培养28天，得到发酵液，所述的发酵培养基每升是通过以下方法配制的：用500mL的纯水煮200g的马铃薯，煮沸20min，过滤得马铃薯汁，再加入葡萄糖20g、海盐30g，用水补足至1000mL。
进一步优选，步骤(b)所述的浓缩是采用减压浓缩。
本发明的第三个目的是提供化合物Penicimeroterpenoids A、B、C或其药用盐在制备胰蛋白酶抑制剂、人白细胞弹性蛋白酶抑制剂、蛋白酪氨酸磷酸酶CDC25B抑制剂、蛋白酪氨酸磷酸酶SHP1抑制剂中的应用。
本发明的第四个目的是提供一种胰蛋白酶抑制剂、人白细胞弹性蛋白酶抑制剂、蛋白酪氨酸磷酸酶CDC25B抑制剂、蛋白酪氨酸磷酸酶SHP1抑制剂，所述的四种酶抑制剂含有化合物Penicimeroterpenoids A、B、C中任一化合物或其药用盐作为活性成分。
本发明的第五个目的是提供真菌Penicilliumsp.SCSIO 41512在制备化合物Penicimeroterpenoids A、B、C中的应用。
本发明从一株海洋真菌Penicilliumsp.SCSIO 41512的发酵产物中分离得到新的3个具有抑制胰蛋白酶、人白细胞弹性蛋白酶、蛋白酪氨酸磷酸酶CDC25B、蛋白酪氨酸磷酸酶SHP1活性的化合物Penicimeroterpenoids A、B、C，它们可用于胰蛋白酶抑制剂、人白细胞弹性蛋白酶抑制剂、蛋白酪氨酸磷酸酶CDC25B抑制剂、蛋白酪氨酸磷酸酶SHP1抑制剂先导化合物的研究。
本发明的真菌Penicilliumsp.SCSIO 41512，于2020年5月22日保藏于广东省微生物菌种保藏中心(GDMCC)，地址：广州市先烈中路100号大院59号楼5楼，其保藏编号为GDMCCNo：61031。
附图说明
图1是化合物1(化合物Penicimeroterpenoid A)的关键HMBC,COSY相关；
图2为化合物1关键NOESY相关；
图3是化合物1(化合物Penicimeroterpenoid A)的单晶衍射结构。
图4为化合物2和3(化合物PenicimeroterpenoidsB and C)的实测CD与理论计算ECD光谱的对比。
具体实施方式
以下实施例是对本发明的进一步说明，而不是对本发明的限制。
实施例1
(1)发酵培养基每升是通过以下方法配制的：用500mL的纯水煮200g的马铃薯，煮沸20min，过滤得马铃薯汁，再加入葡萄糖20g、海盐30g，用水补足至1000mL，在115℃高压蒸汽灭菌25分钟制得。按照此方式配置发酵培养基60L。将60L发酵培养基装入约200个1000mL的三角烧瓶中，每瓶约300mL。
(2)发酵液和菌丝体的制备：将真菌Penicilliumsp.SCSIO 41512在真菌适用的平板培养基中生长，待真菌长出孢子后，用竹签将真菌从平板上移至盛有水的三角瓶中，用移液枪将真菌接种到发酵培养基中(1L三角瓶中盛有300mL发酵培养基)，于室温(26℃)静置培养28天后，收取发酵液和菌丝体，用纱布过滤分开菌液和菌丝体。
(3)化合物分离纯化：将经上述发酵培养基培养得到的菌丝体破碎后用丙酮浸泡，重复提取三次，合并提取液，减压浓缩除去丙酮，剩余水相用乙酸乙酯萃取，浓缩得菌体的乙酸乙酯提取物；将发酵得到的菌液用大孔树脂XAD-16充分吸收，然后用水洗脱去除培养基，再用乙醇或甲醇洗脱，随后减压浓缩得到菌液的提取物；合并菌体和菌液的提取物。将总提取物用正相硅胶(100-200目)干法拌样后，装入玻璃层析柱(H细硅胶)，常温减压柱层析，依次用二氯甲烷：甲醇体积比分别为100:0,95:5,90:10,80:20,70:30,60:40,50:50的二氯甲烷-甲醇系统梯度洗脱，最后洗脱液通过TLC和HPLC检测合并得到10个组分(Fr.1～Fr.10)。
收集二氯甲烷:甲醇体积比95:5冲洗下来的样品得到组分Fr.2；组分Fr.2通过中压反相ODS柱分离，以甲醇/水/TFA(体积比5:95:0.03-100:0:0.03)梯度洗脱，得到11个组分(Fr.2-1–Fr.2-11)；其中用甲醇/水/TFA(v/v/v,65:35:0.03)梯度洗脱得到的组分Fr.2-7经过凝胶Sephadex LH-20柱分离(以甲醇为洗脱溶剂)，通过薄层层析点板，用二氯甲烷：甲醇＝20:1为展开剂，将其中Rf值有明显区别的点(分别为约0.3和0.4)分别合并为组分Fr.2-7-2(Rf＝0.3)和Fr.2-7-1(Rf＝0.4)。Fr.2-7-1进一步通过半制备型HPLC(YMC-PackODS-A column,250×10,5μm,12nm)纯化，以甲醇/水/TFA(v/v/v,70:30:0.03)等度洗脱得到化合物Penicimeroterpenoid A(1)(5.12mg,tR＝10.4min，5mL/min)。Fr.2-7-2通过半制备型HPLC(YMC-Pack ODS-A column,250×10,5μm,12nm)纯化，以甲醇/水/TFA(v/v/v,65:35:0.03)等度洗脱得到化合物PenicimeroterpenoidsB(2)(3.03mg,tR＝17.3min，3mL/min)和C(3)(5.18mg,tR＝18.4min，3mL/min)。
结构推定：
化合物1：无色晶体，高分辨质谱(HRESIMS)在m/z539.2273给出[M+Na]+离子峰，结合13CNMR数据(表1)，推测其分子式为C28H36O9，不饱和度为11。氢谱上的信号显示含有1个甲氧基[δH3.74(s,H3-28)]，1个乙酰化甲基[δH2.00(s,H3-27)]，和其它6个甲基[δH1.83(s,H3-21)，1.75(s,H3-20)，1.42(s,H3-18)，1.21(s,H3-22)，0.94(s,H3-24)，0.85(s,H3-25)]。13CNMR和DEPT谱显示了28个碳信号，其中包括8个甲基，3个亚甲基，5个次甲基(其中2个被氧化)，5个脂肪族季碳(其中1个被氧化)，2个烯烃基非质子化碳，3个羧基，和2个酮羰基。该化合物的核磁数据与citreohybriddiones A和B、andrastins A–D，以及citreohybridonol的相似，推测化合物1也是具有andrastin骨架的混源萜。
通过对二维核磁数据的分析，证实了上述推测。1H-1H COSY图谱(图1)显示了H-2与H-1/H-3相关，结合HMBC图谱显示H-2与C-10(δC45.8)，H-3与C-4/C-5/C-26(δC172.1)，H-5与C-1(δC22.7)/C-3(δC77.2)/C-4(δC35.8)，以及H3-27与C-26相关，表明有1个乙酰基连接在A环的C-3位置上。HMBC数据的H3-24和H3-25与C-4(δC35.8)相关，证实2个甲基连接在C-4位置上。图1中B环主要是通过1H-1H COSY谱显示H-6与H-5/H-7相关、HMBC谱显示H-5与C-10(δC45.8)/C-23(δC180.6)相关、以及H-6与C-10/C-23相关来确定的。进一步分析HMBC图谱，发现H-5与C-9/C-10相关、H-6与C-8/C-10相关、H-7与C-8/C-9相关，确定C环的存在。另外，1H-1H COSY谱显示H-9与H-11相关，HMBC谱显示H-9与C-8/C-12/C-14相关、H-11与C-12/C-13相关、H3-20与C-12/C-13/C-14相关、H3-21与C-11/C-12/C-13相关、H3-22与C-7/C-8/C-9/C-14/C-19(弱)相关，表明D环的存在。此外，通过HMBC谱中H3-28与C-19和C-14相关，表明有一个甲酰基连接在C-14位置上。HMBC图谱显示H3-20与C-14的三键相关，H3-20与C-15(δC207.1)的四键“W”型相关，以及H3-21与C-15的五键相关，表明C-15连接在C-14上。进一步分析HMBC图谱，发现H3-18与C-15/C-16/C-17相关、H-11与C-12/C-13/C-16/C-17相关、H3-21与C-11/C-12/C-13/C-17相关，确定了E环的存在。因此，化合物1的平面结构式(1)所示。
通过对NOESY图谱的分析(图2)，进一步确定了化合物1的相对构型。NOESY图谱显示H-1b与H-5相关、H-2b与H3-24相关、H-3与H3-25相关、以及H-5与H3-24相关，表明H-3是α构型、H-5是β构型。根据NOESY谱中H-5与H-6/H-9相关、H-6与H-7b相关、H-7a与H3-22相关、以及H-7b与H-9相关，表明H-6和H-9是β构型、CH3-22是α构型。基于H-6的构型，确定了A/B/C三环骨架中C-10的构型。H3-18与H3-28间的NOE相关表明它们在E环的同侧。最后，根据H-11与H3-22的NOE相关、以及以C-11和C-14作为桥头碳的双环[3.2.2]壬烷支架的存在，确定了手性碳C-11和C-14的构型。以上构型推测结论被进一步通过分析化合物1的X-ray衍射数据(图3)所证实，最终确定化合物1的绝对构型为3S,5R,6S,8S,9R,10R,11R,14R,16R，命名为Penicimeroterpenoid A。量子化学ECD谱计算进一步证实了化合物1的绝对构型。与其它DMOA衍生物混源萜不同，环D和环E在化合物1中形成了一个新颖的桥环骨架，从而产生了6/5/6/6/7多环体系。
化合物2：淡黄色油状物，高分辨质谱(HRESIMS)在m/z511.2300给出[M+Na]+离子峰，结合13CNMR数据(表1)，推测其分子式为C27H36O8，不饱和度为10。将化合物2的1H和13C的NMR数据(表1)与1和citreohybriddione A的NMR数据比较，发现它们很相似。通过对化合物2二维NMR谱数据(HSQC和HMBC)的详细分析，发现它与citreohybriddione A具有相同的A-D环状片段，它们之间的差异在于2的E环是新颖的四元环，而非五元环。HMBC图谱显示H3-18与C-13/C-15(δC209.7)/C-16(δC87.1)相关、以及H3-20与C-13/C-14/C-16相关，确定C-17上的羰基被降解，从而形成四元环。化合物2的NOESY谱和化合物1的NOESY谱所显示的NOE相关信号基本一致，从而推测化合物2的A-C环中手性中心的相对构型与化合物1的相同。此外，结合NOESY图谱中H-9与H3-18相关、H3-20与H3-22/H3-28相关，表明CH3-20和OCH3-28是α构型、CH3-18是β构型。最后，通过量子化学ECD计算确定了化合物2的绝对构型(图4)。计算构型(3S,5R,6S,8S,9R,10R,13R,14R,16S)-2所得到的加权ECD谱与实验CD谱吻合度高，从而确定了化合物2的绝对构型，命名为PenicimeroterpenoidB。
化合物3：淡黄色油状物，高分辨质谱(HRESIMS)在m/z511.2306给出[M+Na]+离子峰，结合13CNMR数据(表1)，推测其和化合物2有相同的分子式C27H36O8。通过比较化合物2和3的一维和二维NMR数据，推测它们具有相同的碳骨架，区别只在E环上，化合物3的E环是通过C-15位置上的羰基降解而形成的四元环。HMBC谱显示H3-18与C-14/C-16(δC86.5)/C-17(δC218.8)相关、H3-20与C-12/C-13/C-14/C-17相关，证实了化合物3中E环的存在。通过分析比较化合物3和化合物2的NOESY谱所显示的NOE相关信号，发现除了C-16的构型外，化合物3中手性中心的构型与2的构型相同，其中H3-28(δH3.65)与H3-18(δH1.43)相关、H3-20与H3-22/H3-28相关，表明CH3-18是α构型，从而确定了C-16的相对构型。化合物3的绝对构型也是通过量子化学ECD计算确定的(图3)，计算结果表明，加权的(3S,5R,6S,8S,9R,10R,13R,14R,16S)-3的ECD谱与实验CD谱吻合度高，从而确定了化合物3的绝对构型为3S,5R,6S,8S,9R,10R,13R,14R,16S，命名为PenicimeroterpenoidC。
表1.1H(700MHz)and 13C NMR(175MHz)data of 1–3inmethanol-d4(δin ppm,J inHz).
实施例2化合物Penicimeroterpenoids A-C的蛋白酪氨酸磷酸酶活性测试
将人蛋白酪氨酸磷酸酶CDC25B、SHP2、MEG2、SHP1、TCPTP、CD45或PTP1B的基因插入表达载体中，然后克隆到大肠杆菌(Escherichia coli)中，再进行表达纯化获得人蛋白酪氨酸磷酸酶CDC25B、SHP2、MEG2、SHP1、TCPTP、CD45或PTP1B。酶抑制活性的测定是在96孔板中使用对硝基苯磷酸(pNPP)作为底物，每孔板含100μL的反应混合物。分别将人类重组蛋白酪氨酸磷酸酶CDC25B、SHP2、MEG2、SHP1、TCPTP、CD45或PTP1B(0.05μg)添加到50μL含有50mMHEPES，100mMNaCl，1mM EDTA及1mMdithiothreitol(DTT)的反应缓冲液中(pH6.5)，再在每个96孔板中分别添加待测的化合物样品(化合物Penicimeroterpenoids A-C)。Na3VO4作为阳性对照，DMSO作为阴性对照，用于评价此高通量筛选系统。在室温下预孵育15分钟后，添加50μL含有50mMpNPP的缓冲液，并继续在37℃下培养60分钟。磷酸酶活性是通过测定所产生的对硝基苯酚在405nm处的吸光度来测定的。IC50值是使用Gen5软件计算(Synergy2Multi-Mode Microplate Reader,BioTek Instruments,Inc.,headquartered inWinooski,VT,USA)。每个实验重复3次。
实施例3化合物Penicimeroterpenoids A-C的胰蛋白酶和人白细胞弹性蛋白酶活性测试
抑制人白细胞弹性蛋白酶(HLE)的具体方法参考(Zheng,Z.；Zhang,S.；Lu,X.；Ma,Y.；Fan,Y.；Shi,Y.；Dong,A.；Duan,B.Biol.Pharm.Bull.2012,35,2247–2251.)所述。简述如下：50μl反应缓冲液(N-(2-hydroxyethyl)piperazine-N’-2-ethanesulfonic acid(HEPES)pH 6.5,0.5M NaCl)含有0.0015单元HLE和2μL被测试样品(溶解在DMSO中)或DMSO(空白对照)在96孔板中室温孵化15分钟，反应开始时，加入50μl底物溶液(含有最终浓度为300μMN-MeO-Suc-Ala-Ala-Pro-Val-pNA)。在37℃孵化30分钟后，使用微型平板阅读器测定对硝基苯胺的水解产物在405nm下的OD值。被测试样品的浓度是40μg/mL。每个实验重复3次。
抑制胰蛋白酶(trypsin)的实验方法和上述HLE酶测试方法相同，只是底物改用300μMBz-Ile-Glu-Gly-Arg-pNA、缓冲液改用含0.15M NaCl的0.05M Tris–HCl(pH7.2)。
实验结果如表2所示，上述蛋白酪氨酸磷酸酶、胰蛋白酶和人白细胞弹性蛋白酶活性测试结果显示：化合物penicimeroterpenoids A–C有较温和的抑制胰蛋白酶、人白细胞弹性蛋白酶、蛋白酪氨酸磷酸酶CDC25B、蛋白酪氨酸磷酸酶SHP1的活性。
表2化合物1-3对9种酶的抑制活性
以上仅是本发明的优选实施方式，应当指出的是，上述优选实施方式不应视为对本发明的限制，本发明的保护范围应当以权利要求所限定的范围为准。对于本技术领域的普通技术人员来说，在不脱离本发明的精神和范围内，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (6)
1.化合物Penicimeroterpenoids A、B、C或其药用盐，其结构式如式（Ⅰ）所示：
式（Ⅰ）。
2.一种权利要求1所述的化合物Penicimeroterpenoids A、B、C的制备方法，其特征在于，包括以下步骤：
（a）制备真菌Penicilliumsp.SCSIO41512的发酵产物，发酵产物分离出发酵液和菌丝体；
（b）将步骤（a）得到的发酵液用大孔树脂吸附，接着用水冲洗大孔树脂去掉培养基成分，再用甲醇或乙醇冲洗大孔树脂，浓缩有机相得到甲醇或乙醇提取物；或将步骤（a）得到的发酵液用乙酸乙酯、二氯甲烷或氯仿溶剂萃取，浓缩得到乙酸乙酯提取物、二氯甲烷提取物或氯仿提取物；菌丝体破碎后用丙酮浸泡，提取液减压浓缩除去丙酮，剩余水相用乙酸乙酯、二氯甲烷或氯仿萃取，浓缩得菌丝体的乙酸乙酯、二氯甲烷或氯仿提取物；合并菌丝体和发酵液的提取物，得到总提取物；
（c）将步骤（b）的总提取物经过正相硅胶柱层析，依次用二氯甲烷:甲醇体积比为100:0, 95:5, 90:10, 80:20, 70:30, 60:40, 50:50的溶剂系统梯度洗脱，收集二氯甲烷:甲醇体积比95:5冲洗下来的样品得到组分Fr.2；组分Fr.2通过中压反相ODS柱分离，以甲醇/水/TFA体积比5:95:0.03-100:0:0.03梯度洗脱，其中用甲醇/水/TFA 按体积比65:35:0.03梯度洗脱得到的组分Fr.2-7经过凝胶Sephadex LH-20柱分离，以甲醇为洗脱溶剂，结合TLC，以二氯甲烷：甲醇=v/v 20:1为展开剂，分别收集Rf为0.3的流份为组分Fr.2-7-2，Rf为0.4的流份为Fr.2-7-1，Fr.2-7-1进一步通过半制备型HPLC纯化，以甲醇/水/TFA按体积比70:30:0.03等度洗脱得到化合物Penicimeroterpenoid A，Fr.2-7-2通过半制备型HPLC纯化，以甲醇/水/TFA按体积比65: 35: 0.03等度洗脱得到化合物PenicimeroterpenoidsB和C；
步骤（a）中所述的发酵液和菌丝体是通过以下方法制备：将真菌Penicilliumsp.SCSIO41509在真菌适用的平板培养基中生长，待真菌长出孢子后，将真菌接种到发酵培养基中，于室温静置培养28天，得到发酵液和菌丝体，所述的发酵培养基每升是通过以下方法配制的：用500 mL的纯水煮200 g的马铃薯，煮沸20 min，过滤得马铃薯汁，再加入葡萄糖20 g、海盐30 g，用水补足至1000 mL；
所述的真菌Penicilliumsp.SCSIO 41512，于2020年5月22日保藏于广东省微生物菌种保藏中心，地址：广州市先烈中路100号大院59号楼5楼，其保藏编号为GDMCC No：61031。
3.根据权利要求2所述的制备方法，其特征在于，步骤（b）中所述的浓缩是采用减压浓缩。
4. 权利要求1所述的化合物Penicimeroterpenoids A、B、C或其药用盐在制备胰蛋白酶抑制剂、人白细胞弹性蛋白酶抑制剂、蛋白酪氨酸磷酸酶CDC25B抑制剂和/或SHP1酶抑制剂中的应用。
5. 一种胰蛋白酶抑制剂、人白细胞弹性蛋白酶抑制剂、蛋白酪氨酸磷酸酶CDC25B抑制剂和/或蛋白酪氨酸磷酸酶SHP1抑制剂，其特征在于，含有权利要求1所述的化合物Penicimeroterpenoids A、B、C中任一化合物或其药用盐作为活性成分。
6. 真菌Penicilliumsp.SCSIO 41512在制备权利要求1中的化合物Penicimeroterpenoids A、B和/或C中的应用，所述的真菌Penicilliumsp.SCSIO 41512，于2020年5月22日保藏于广东省微生物菌种保藏中心，地址：广州市先烈中路100号大院59号楼5楼，其保藏编号为GDMCC No：61031。
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