A New Strategy for Deleting Animal drugs from Traditional Chinese Medicines based on Modified Yimusake Formula

Abstract

Traditional Chinese medicine (TCM), such as Uyghur Medicine (UM) has been used in clinical treatment for many years. TCM is featured as multiple targets and complex mechanisms of action, which is normally a combination of medicinal herbs and sometimes even contains certain rare animal medicinal ingredients. A question arises as to whether these animal materials can be removed replaced from TCM applications due to their valuable rare resources or animal ethics. Here, we select a classical UM Yimusake formula, which contains 3 animal drugs and other 8 herbs, and has got wealthy experience and remarkable achievements in treating erectile dysfunction (ED) in China. The active components, drug targets and therapeutic mechanisms have been comprehensively analyzed by systems-pharmacology methods. Additionally, to validate the inhibitory effects of all candidate compounds on their related targets, in vitro experiments, computational analysis and molecular dynamics simulations were performed. The results show that the modified, original and three animal materials display very similar mechanisms for an effective treatment of ED, indicating that it is quite possible to remove these three animal drugs from the original formula while still keep its efficiency. This work provides a new attempt for deleting animal materials from TCM, which should be important for optimization of traditional medicines.

Figure 1

C-Tv network. 11 bioactive compounds (cyan squares) from Myristica Semena, 8 ones from Stigma Croci (dark orange squares) and 11 ones from Gymnadenia conopsea (yellow green squares) predicted to have 124 potential protein targets (circles). The pink circles (53) are the common targets of three herbs. The orange (27) and green (1) circles are the specific targets of Stigma Croci and Gymnadenia conopsea, respectively. Node size is proportional to its degree.

Figure 2

The global view of C-Tc network for Semen Strychni and Myristica Semena which is soly related to the CNS diseases. The net is composed of 11 bioactive compounds (cyan squares) from Myristica Semena and 12 ones from Stigma Croci (orange squares), as well as 78 potential protein targets (circles) these compounds interact with. The pink circles (53) are the common targets of both herbs. Node size is proportional to its degree.

Figure 3

C-Tv network. The net is composed of 7 bioactive compounds from Syringa oblata (cyan squares), 9 ones from Boswellia (orange squares), 15 ones from Rhizoma Alpiniae Officinarum (olive squares) and 11 ones from Gymnadenia conopsea (red squares) as well as the 132 potential protein targets (circles) that these compounds interact with. The pink circles (53) are the common targets of four herbs. The purple circles are the specific targets of Syringa oblata. Node size is proportional to its degree.

Figure 4

Computational modeling of F2 with their ligands. (A) F2-curcumin, (B) F2-galangin, (C) F2-quercetin, (D) F2-methyllinolenate, (E) F2-macelignan, (F) F2-isorhamnetin, (G) F2-kaempferol. The molecules are displayed as a ball and stick model, H-bonds are shown as dotted black lines, with distance unit of Å. Other atoms O and N are colored as red and blue, respectively.

Figure 5

Computational modeling of MABO with their ligands. (A) MAOB-chrysin, (B) MAOB-eugenol, (C) MAOB-kaempferol, (D) MAOB-macelignan. The molecules are displayed as a ball and stick model, H-bonds are shown as dotted black lines, with distance unit of Å. Other atoms O and N are colored as red and blue, respectively.

Figure 6

RMSD profiles of the ligands in complexes with F2 (A) and MAOB (B) for the backbone from the starting structures during the equilibration process.

Figure 7

Hydrogen-bonding networks at the active site of the ligand-F2 complex from snapshots of the conformations obtained in 5-ns MD simulation for compounds (A) curcumin, (B) methyllinolenate, (C) macelignan, (D) isorhamnetin, (E) kaempferol, (F) quercetin, (G) galangin. The molecules are displayed as ball and stick models. H-bonds are shown as dotted black lines, with distance unit of Å. Other atoms O and N are colored as red and blue, respectively.

Figure 8

Binding conformations of different compounds in MAOB after the MD simulation. (A) chrysin, (B) eugenol, (C) kaempferol, (D) macelignan.

Figure 9

The C-T-P network was constructed by overlaying the C-T network onto T-P network. The squares and circles, respectively, represent potential compounds and targets. The pathway node is represented as red hexagon.

Figure 10

Distribution of target proteins of modified Yimusake formula on the compressed ED pathway.

Figure 11

C-T network. 32 compounds (orange squares) from the three animal materials Moschus, Ambra Grisea and Bullwhip and 66 (cyan squares) ones from the modified Yimusake formula are connected with 139 potential protein targets (circles). The pink circles are the common targets of the three animal materials and the modified Yimusake formula. The blue circles are the specific targets of animal materials, which are not associated with ED.

Figure 12

C-T network. 66 compounds (orange squares) from the modified Yimusake formula and 81 ones from original Yimusake formula (green squares) are connected with their potential protein targets (circles). The blue circles are the common targets of both formulae, while the red circles are the specific targets of original Yimusake formula, which are not associated with ED.