Muscle-type creatine kinase (CK-MM) is the target protein of ginsenosides in skeletal muscle. 20(S)-protopanaxadiol [20(S)-PPD] is an activator of CK-MM and exerts an anti-fatigue effect. In this study, twelve dammarane-type compounds were used for structure-activity relationship analysis in terms of enzyme activity, intermolecular interaction, and molecular docking. Enzyme activity analysis showed that 20(S)-PPD, 20(R)-PPD, 20(S)-protopanaxatriol [20(S)-PPT], 25-OH-PPD, 24-COOH-PPD, panaxadiol (PD), and ginsenoside Rh2 significantly increased CK-MM activity. Panaxatriol (PT), ocotillol, ginsenoside Rg1, and ginsenoside Rd had no significant influence on CK-MM activity, while jujubogenin inhibited its activity. Biolayer Interferometry (BLI) assay produced the same results as those on enzyme activity. The interaction intensity between dammarane-type compounds and CK-MM was linearly related to the compounds’ maximum increment rate of enzyme activity. Molecular docking showed the following sequence of docking scores: Rd > Rg1 > Rh2 > 24-COOH-PPD > 20(S)-PPD > 20(S)-PPT > 25-OH-PPD > 20(R)-PPD > ocotillol > PT > PD > jujubogenin. We demonstrated that 20(S)-PPD was the best activator of CK-MM among the 12 dammarane-type compounds. The cyclization of the dammarane side chain, the hydroxyl group at position C⁶, and the glycosylation of C³, C⁶, and C²⁰ reduced the ability to activate CK-MM. These findings can help in the development of enhanced CK-MM activators through structural modification.

肌肉型肌酸激酶（CK-MM）是骨骼肌中人参皂甙的靶蛋白。20（S）-原人参二醇[20（S）-PPD]是CK-MM的活化剂，具有抗疲劳作用。在这项研究中，从酶活性，分子间相互作用和分子对接方面，使用了十二种丹玛烷型化合物进行构效关系分析。酶活性分析表明20（S）-PPD，20（R）-PPD，20（S）-原托那三醇[20（S）-PPT]，25-OH-PPD，24-COOH-PPD，人参二醇（PD）和人参皂苷Rh2显着增加CK-MM活性。人参三醇（PT），烟碱，人参皂苷Rg1和人参皂苷Rd对CK-MM活性无显着影响，而大枣生成素则抑制其活性。生物层干涉测定法（BLI）的测定结果与酶活性相同。达玛烷型化合物与CK-MM的相互作用强度与化合物的最大酶活性增加率呈线性关系。分子对接显示了对接分数的以下顺序：Rd> Rg1> Rh2> 24-COOH-PPD> 20（S）-PPD> 20（S）-PPT> 25-OH-PPD> 20（R）-PPD> ocotillol > PT> PD> jujubogenin。我们证明了20（S）-PPD是12种达玛烷类型化合物中CK-MM的最佳活化剂。达玛烷侧链的环化，C位置的羟基 大枣素。我们证明了20（S）-PPD是12种达玛烷类型化合物中CK-MM的最佳活化剂。达玛烷侧链的环化，C位置的羟基 大枣素。我们证明了20（S）-PPD是12种达玛烷类型化合物中CK-MM的最佳活化剂。达玛烷侧链的环化，C位置的羟基^{如图6所示}，C ³，C ⁶和C ²⁰的糖基化降低了激活CK-MM的能力。这些发现可通过结构修饰帮助开发增强型CK-MM活化剂。