Phenylspirodrimane with Moderate Reversal Effect of Multidrug Resistance Isolated from the Deep-Sea Fungus Stachybotrys sp. 3A00409

Two new phenylspirodrimanes, stachybotrins K and L (1 and 2), together with eight known analogues (3–10), were isolated from deep-sea-derived Stachybotrys sp. MCCC 3A00409. Their structures were determined by extensive NMR data and mass spectroscopic analysis. Absolute configurations of new compounds were determined through a comparison of their circular dichroism (CD) spectra with other reported compounds. The possible reversal effects of all compounds were assayed in the resistant cancer cell lines. Stachybotrysin B (8) can reverse multidrug resistance (MDR) in ABCB1-overexpression cells (KBv200, Hela/VCR) at the non-cytotoxic concentration. Doxorubicin accumulation assay and molecular-docking analysis reveal that the mechanism of its reversal MDR effect may be related to the increase in the intracellular concentration of substrate anticancer drugs.


Introduction
Chemotherapy is the main treatment method for many malignant tumors.However, drug resistance is becoming a major problem in several cancers.Multidrug resistance (MDR) refers to the resistance of tumor cells to multiple chemotherapy agents, mediated by drug inactivation or the removal of drugs from target tumor cells [1,2].MDR is currently considered to be one of the most common causes of cancer chemotherapy failure [3].The most common mechanism of MDR is the overexpression of ABC transporters, which actively pump out large amounts of chemotherapy drugs from cancer cells, thereby weakening the efficacy of chemotherapy drugs [4,5].The subfamily ABCB1/MDR1/P-glycoprotein, ABCC1/MRP1, and ABCG2/BCRP are the most extensively studied and considered as prime factors for the induction of MDR in tumor cells [6,7].Among them, ABCB1/P-glycoprotein (P-gp) remains the best-studied and most potent ABC transporter to induce chemoresistance, and the upregulation of ABCB1/P-gp is closely linked with the emergence of MDR in tumor cells [8,9].The discovery of ABCB1/P-gp suggests that combining ABCB1/P-gp inhibitors with traditional anticancer drugs may be a promising strategy for surmounting P-gp-mediated MDR [10,11].In recent decades, three generations of ABCB1/P-gp inhibitors have been developed, some of which are currently undergoing clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.

P-gp inhibitor Mucositis and neutropenic fever
Phase II Zosuquidar (LY335979) clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.
clinical trials to evaluate their role in circumventing anti-cancer resistance [9,[12][13][14][15][16] (see Table 1).However, due to their unacceptable toxicity and problematic pharmacokinetic interactions, many of these inhibitors have not been used in clinical practice [8,17,18].Therefore, there is still a strong demand for the screening of multidrug-resistance reversal agents and the development of new multidrug-resistance antagonists.ONT-093 P-gp inhibitor Not significant Phase I Stachybotrys, a genus of filamentous fungi, is known to produce a broad range of secondary metabolites, such as macrocyclic trichothecenes, atranones, and phenylspirodrimanes (PSDs) [19,20].PSDs belong to a type of meroterpenoids, which are proposed to be derived from the hybrid polyketide-terpenoid biosynthetic pathway.They feature a common drimane-type sesquiterpene skeleton connected with a benzene ring Stachybotrys, a genus of filamentous fungi, is known to produce a broad range of secondary metabolites, such as macrocyclic trichothecenes, atranones, and phenylspirodrimanes (PSDs) [19,20].PSDs belong to a type of meroterpenoids, which are proposed to be derived from the hybrid polyketide-terpenoid biosynthetic pathway.They feature a common drimane-type sesquiterpene skeleton connected with a benzene ring through a spirofuran ring and show high structural diversity [19].To date, over 120 compounds of this class, including monomeric and dimeric PSDs, have been discovered to derive from Stachybotrys [19][20][21][22][23].These metabolites are designated as the most dominant group of mycotoxins in this genus [19,20,24].PSDs can be further divided into three main classes: tetracyclic aromatic sesquiterpenoids with alcohol and/or aldehyde side chains, like stachybotrydial, pentacyclic aromatic sesquiterpenoids, such as stachybotrylactams and stachybotrylactones, and stachyflins with pentacyclic moiety including a cis-fused decalin [22,25].They display diverse biological activities, including anticomplement, antiviral (HIV-1 and IAV) and anti-inflammatory activity, cytotoxicity, neuroprotective effects, tyrosine kinase inhibition, and tumor-related kinases inhibition [19].Active monomeric compounds are mostly concentrated in the first class with alcohol and/or aldehyde functionalities and stachybotrylactones [19].

ONT-093
P-gp inhibitor Not significant Phase I Stachybotrys, a genus of filamentous fungi, is known to produce a broad range of secondary metabolites, such as macrocyclic trichothecenes, atranones, and phenylspirodrimanes (PSDs) [19,20].PSDs belong to a type of meroterpenoids, which are proposed to be derived from the hybrid polyketide-terpenoid biosynthetic pathway.They feature a common drimane-type sesquiterpene skeleton connected with a benzene ring through a spirofuran ring and show high structural diversity [19].To date, over 120 compounds of this class, including monomeric and dimeric PSDs, have been discovered to derive from Stachybotrys [19][20][21][22][23].These metabolites are designated as the most dominant group of mycotoxins in this genus [19,20,24].PSDs can be further divided into three main classes: tetracyclic aromatic sesquiterpenoids with alcohol and/or aldehyde side chains, like stachybotrydial, pentacyclic aromatic sesquiterpenoids, such as stachybotrylactams and stachybotrylactones, and stachyflins with pentacyclic moiety including a cis-fused decalin [22,25].They display diverse biological activities, including anticomplement, antiviral (HIV-1 and IAV) and anti-inflammatory activity, cytotoxicity, neuroprotective effects, tyrosine kinase inhibition, and tumor-related kinases inhibition [19].Active monomeric compounds are mostly concentrated in the first class with alcohol and/or aldehyde functionalities and stachybotrylactones [19].

8′
48.  Compound 2 was obtained as a pale yellow oil and was assigned the molec mula of C30H37NO5 on the basis of HRESIMS at m/z 514.2570 [M + Na] + (ca 514.2569), along with 1 H and 13 C NMR spectroscopic data (Table 2).The structure determined as a methylated analogue of 1 on the basis of the close similarity of NM except for the presence of the methoxyl resonances at δH 3.79 (3H, s) and δC 55.6, w methoxy group was substituted at OH-10′, as evident from the correlation betwee (δH 3.79) and C-10′ (δC 154.9) (Supplementary Materials: Figure S1).The relative ration of the phenylspirodrimane moiety in 2 was further established by its NOE relations (Figure S1).According to the similar chemical shifts from 2 to 1 (Figur   Compound 2 was obtained as a pale yellow oil and was assigned the molecular formula of C30H37NO5 on the basis of HRESIMS at m/z 514.2570 [M + Na] + (calcd.For 514.2569), along with 1 H and 13 C NMR spectroscopic data (Table 2).The structure of 2 was determined as a methylated analogue of 1 on the basis of the close similarity of NMR data, except for the presence of the methoxyl resonances at δH 3.79 (3H, s) and δC 55.6, while the methoxy group was substituted at OH-10′, as evident from the correlation between H3-15′ (δH 3.79) and C-10′ (δC 154.9) (Supplementary Materials: Figure S1).The relative configuration of the phenylspirodrimane moiety in 2 was further established by its NOESY correlations (Figure S1).According to the similar chemical shifts from 2 to 1 (Figure 3) and Stachybotrin H (4) was previously isolated from the same strain [26], but the absolute configuration was not established.Through a comparison of its ECD curves with 1 and 2 (Figure 3), the absolute configuration of 4 was also established as 3R, 5S, 8R, 9R, and 10S.

Biological Assays
To quickly screen for compounds with resistance reversal activity, we used flow cytometry analysis to detect the effects of compounds 1-10 on the accumulation of doxorubicin (DOX) in the resistant human oral epithelial cancer cell line KBv200.The results showed that compound 8 could increase the accumulation of DOX in KBv200 cells at 20 µM (Figure 4).
To quickly screen for compounds with resistance reversal activity, we used fl tometry analysis to detect the effects of compounds 1-10 on the accumulation of d bicin (DOX) in the resistant human oral epithelial cancer cell line KBv200.The showed that compound 8 could increase the accumulation of DOX in KBv200 cel µM (Figure 4).To find the suitable concentrations of compound 8 for reversing MDR in vi first examined the cytotoxic effect of 8 on different cancer cell lines via MTT ass shown in Figure 5, stachybotrysin B (8) had almost no significant cytotoxic effect KBv200 cells at concentrations below 10 µM, and had no cytotoxic effects on He Hela/VCR below 80 µM, after 72 h of treatment.In addition, compound 8 showed totoxic activity against normal H9C2 cells below 40 µM (Figure 5E).Thus, we cho centrations of 10 µM and 20 µM as the maximum concentrations for further reve says in KBv200 and Hela/VCR cell lines, respectively; at the chosen concentrations than 90% of cells survived.To find the suitable concentrations of compound 8 for reversing MDR in vitro, we first examined the cytotoxic effect of 8 on different cancer cell lines via MTT assay.As shown in Figure 5, stachybotrysin B (8) had almost no significant cytotoxic effect on KB, KBv200 cells at concentrations below 10 µM, and had no cytotoxic effects on Hela and Hela/VCR below 80 µM, after 72 h of treatment.In addition, compound 8 showed no cytotoxic activity against normal H9C2 cells below 40 µM (Figure 5E).Thus, we chose concentrations of 10 µM and 20 µM as the maximum concentrations for further reversal assays in KBv200 and Hela/VCR cell lines, respectively; at the chosen concentrations, more than 90% of cells survived.
tometry analysis to detect the effects of compounds 1-10 on the accumulation of doxorubicin (DOX) in the resistant human oral epithelial cancer cell line KBv200.The results showed that compound 8 could increase the accumulation of DOX in KBv200 cells at 20 µM (Figure 4).To find the suitable concentrations of compound 8 for reversing MDR in vitro, we first examined the cytotoxic effect of 8 on different cancer cell lines via MTT assay.As shown in Figure 5, stachybotrysin B (8) had almost no significant cytotoxic effect on KB, KBv200 cells at concentrations below 10 µM, and had no cytotoxic effects on Hela and Hela/VCR below 80 µM, after 72 h of treatment.In addition, compound 8 showed no cytotoxic activity against normal H9C2 cells below 40 µM (Figure 5E).Thus, we chose concentrations of 10 µM and 20 µM as the maximum concentrations for further reversal assays in KBv200 and Hela/VCR cell lines, respectively; at the chosen concentrations, more than 90% of cells survived.Then, we investigated the reversal effect of compound 8 on different tumor-sensitive cell lines and their drug-resistant cell lines.The results (Table 3) revealed that compound 8 exhibited moderate resistance reversal effects against the substrate chemotherapeutic agents, DOX and navelbine (NVB), in the ABCB1-overexpressing drug-resistant cell line KBv200, and against DOX in the ABCB1-overexpressing drug-resistant cell line Hela/VCR, with fold resistance ranging from 1.54 to 4.86 and from 1.43 to 7.82, respectively, in a dose-dependent manner.However, compound 8 did not significantly decrease the IC 50 of cisplatin, which was not a substrate chemotherapeutic drug of ABCB1, in the ABCB1overexpressing MDR cells.These results indicated that compound 8 could reverse ABCB1mediated MDR.
We further detected an effect of compound 8 on the intracellular accumulation of DOX, the substrate agents of the ABCB1 transporter, in ABCB1-overexpressing cell lines (KBv200, Hela/VCR) via flow cytometry analysis.The results showed that compound 8 could increase the accumulation of DOX in KBv200 and Hela/VCR, thereby increasing the concentration of DOX in these cancer cell lines at concentrations below 10 µM and 20 µM, respectively (Figure 6).We further detected an effect of compound 8 on the intracellular accumulation of DOX, the substrate agents of the ABCB1 transporter, in ABCB1-overexpressing cell lines (KBv200, Hela/VCR) via flow cytometry analysis.The results showed that compound 8 could increase the accumulation of DOX in KBv200 and Hela/VCR, thereby increasing the concentration of DOX in these cancer cell lines at concentrations below 10 µM and 20 µM, respectively (Figure 6).

Docking Analysis of Compound 8 and Verapamil with ABCB1
Molecule docking simulation technology is a convenient and effective method to explore the interaction between small molecules and targets [32][33][34][35].We performed a molecular docking simulation to characterize the molecular basis of the interactions between compound 8 and the human homology model of the ABCB1 transporter protein structure (PDB ID: 6QEX).The results (Figure 7) revealed that 8 was well-fitted at the substrate-binding site in the transmembrane domains (TMDs), with the highest docking score of −7.4 kcal/mol.Then, we chose this protomol to dock verapamil (a classical ABCB1/P-gp inhibitor) [9] to TMDs.The docking score was −7.7 kcal/mol.rapamil, compound 8 also interact with Gln725 of ABCB1 through H-bonds (Figur In addition, a hydrogen bond was found between the hydroxyl and aldehyde grou the aromatic ring of and ALA-987 and ASN-721.The molecule was also predicted teract with PHE-983, ALA-987, VAL-991, and PHE-336 via hydrophobic interaction.we can infer that the ABCB1 protein might recognize both 8 and verapamil thro similar mechanism.

General Experimental Procedure
Optical rotations were performed using a JASCO P-1020 digital polarimeter Corporation, Tokyo, Japan).ECD spectra were obtained with a JASCO J-815 spectrop imeter (Jasco Corporation, Tokyo, Japan).UV spectra were obtained using a Shimad 210A spectrometer (Shimadzu Corporation, Tokyo, Japan).NMR spectra were rec with 400 and 600 MHz Bruker Avance NMR spectrometers (Bruker BioSpin Fällanden, Switzerland).ESIMS was performed using a Shimadzu LCMS-8040 L Chromatograph Mass Spectrometer (Shimadzu Corporation, Tokyo, Japan).HRE spectra were obtained using a Thermo Scientific Ultimate 3000 UHPLC-Q Exactive trometer (ThermoFisher Scientific, Waltham, MA, USA).Semi-preparative HPLC wa formed on a Waters 1525 system using a semi-preparative C18 (Waters SunFire C18 Prep Column, 10 × 250 mm, 5 μm, 3 mL/min) column coupled with a Waters 2996 p diode array detector (Waters Corporation, Milford, MA, USA).Thin-layer chrom raphy (TLC) was performed on plates precoated with silica gel GF254 (10-40 µm) ( dao Marine Chemical Factory, Qingdao, China).Sephadex LH-20 (Amersham Bioscie For verapamil, a hydrogen bond is formed between the hydroxy group and GLN725 of ABCB1, and hydrophobic interactions are formed between verapamil and five amino acids, like ILE-340, PHE-983, PHE-336, ALA-229, and PHE-343 (Figure 7B).Similar to verapamil, compound 8 also interact with Gln725 of ABCB1 through H-bonds (Figure 7A).In addition, a hydrogen bond was found between the hydroxyl and aldehyde groups at the aromatic ring of and ALA-987 and ASN-721.The molecule was also predicted to interact with PHE-983, ALA-987, VAL-991, and PHE-336 via hydrophobic interaction.Thus, we can infer that the ABCB1 protein might recognize both 8 and verapamil through a similar mechanism.

Fungal Material
The fungus strain Stachybotrys sp.MCCC 3A00409 was isolated from Atlantic Ocean (−2807 m), as described previously [26].The strain was deposited in the Marine Culture Collection Center (MCCC), Third Institute of Oceanography, Ministry of Natural Sources, and School of Pharmacy, Fujian Medical University.

Extraction and Isolation
After incubation, the solid culture was extracted with CH 3 OH three times.The CH 3 OH extract was evaporated under reduced pressure to obtain an aqueous solution, and then the aqueous solution was extracted three times with EtOAc to obtain a brown crude gum (30.2 g).

Cell Cytotoxicity Assay
Cell cytotoxicity assay in vitro was evaluated by MTT assay, as described previously [36].In brief, cells were seeded in 96-well plates at an appropriate density and incubated for 24 h at 37 • C.Then, the cells were treated with various concentrations of chemotherapeutic durgs or compounds.A total of 20 µL of MTT solution (5 mg/mL in PBS) was added to each well after 72 h incubation.The plates were incubated at 37 • C for another 4 h and the resulting formazan crystals were dissolved with 150 µL DMSO.Absorbance in each well was measured by Model 550 Microplate Reader (Bio-Rad, Hercules, CA, USA) at 570 nm.The half maximal (50%) inhibitory concentration (IC 50 ) was calculated from the curves using GraphPad Prism 9.0, using the Bliss method [37].The maximal concentration of compound 8 indicated that a cell survival of more than 80% was required when testing its MDR-reversal effect.The effect of the MDR reversal by compound 8 was calculated by dividing the IC 50 of cells treated with chemotherapeutic drugs alone or by the IC 50 of cells treated with chemotherapeutic drugs in the presence of compound 8.All experiments were repeated at least three times.

DOX Accumulation Assay
A flow cytometry assay was performed to examine whether compounds 1-10 could affect the accumulation of DOX in KBv200 or Hela/VCR cells, as previously described [36].In brief, the logarithmically growing KBv200 or Hela/VCR cells were seeded in six-well plates at a density of 3 × 10 5 per well.After 24 h of incubation, the cells were pretreated with 20 µM of compounds 1-10 or various concentrations of compound 8 or the vehicle for 3 h at 37 • C.Then, cells were further incubated with 10 µM DOX for another 3 h.After that, the cells were collected, washed three times with cold phosphate-buffered saline (PBS), and resuspended in 500 µL PBS.The mean fluorescence intensity (MFI) of DOX was examined via flow cytometry (Cytomics FC500, Beckman Coulter Inc., Brea, CA, USA) with an excitation wavelength of 488 nm and an emission wavelength of 550 nm.

Molecular Docking Analysis
The crystal structures of the ABCB1 protein [38] (PDB ID: 6QEX) used for docking were downloaded from the PDB database.The 3D structure of the small molecule complex was constructed using Chem3D 20.0 and energy was minimized under the MMFF94 force field.Before ligand docking, PyMol 2.5.5 [39] was used to treat the receptor protein, including the removal water molecules, salt ions, and small molecules.Subsequently, a 25 × 25 × 25 grid box, with the center being the center of the crystal ligand, was set up.In addition, ADFRsuite 1.0 [40] was used to convert all processed small molecules and receptor proteins into the necessary PDBQT format for AutoDock Vina 1.1.2[41] docking software.When docking, the global search granularity was set to 32, while the remaining parameters remained at their default settings.The top-scoring model (based on the docking score in kcal/mol) was visualized and analyzed using PyMol 2.5.5.

Data Analysis
All data are expressed as mean ± standard deviation (mean ± SD) established in different experiments and analyzed by one-way analysis of variance (ANOVA), followed by Bonferroni post hoc test.Statistical graphs were produced by GraphPad Prism 9.0.0 software (GraphPad Software, Inc., San Diego, CA, USA).*, p < 0.05; **, p < 0.01; ***, p < 0.001, was considered statistically significant.

Conclusions
In this study, two new phenylspirodrimanes, stachybotrin K (1), and stachybotrin L (2), together with eight known derivatives (3-10), were isolated from deep-sea-derived Stachybotrys sp.MCCC 3A00409.Extensive NMR spectroscopic analysis and ECD analysis were used to elucidate the structures of new compounds, including their absolute configurations.The screening results of all compounds with resistance reversal activity in a drug-resistant tumor cell line showed that stachybotrysin B (8) had the most significant effect.Compound 8 exhibited resistance reversal effects against the substrate chemotherapeutic agents, doxorubicin (DOX), and navelbine (NVB) in the ABCB1-overexpressing drug-resistant cell line KBv200, and against DOX in the ABCB1-overexpressing drug-resistant cell line Hela/VCR, in a dose-dependent manner.The results of flow cytometry analysis showed that the mechanism of its MDR reversal effect might be related to the increase in the intracellular concentration of substrate anticancer drug.The docking analysis of compound 8 binding to ABCB1 transporter protein indicated that 8 can potentially strongly interact with several amino acid residues within the transmembrane regions of ABCB1.This interaction might inhibit the efflux of substrates such as DOX, thereby increasing the accumulation of DOX in ABCB1-resistant cancer cells and reversing ABCB1-mediated multidrug resistance.

Figure 4 .
Figure 4.The effect of compounds 1-10 on the accumulation of DOX in KBv200 cells.** p < p < 0.001 versus control group.

Figure 7 .
Figure 7. Docking of compound 8 (A) and verapamil (B) in the drug-binding pocket of A Compound 8 and verapamil are shown as molecular models via yellow sticks.The light cya toon represents ABCB1.Blue dashed lines represent hydrogen bonding, and gray dashed line resent hydrophobic interactions.

Figure 7 .
Figure 7. Docking of compound 8 (A) and verapamil (B) in the drug-binding pocket of ABCB1.Compound 8 and verapamil are shown as molecular models via yellow sticks.The light cyan cartoon represents ABCB1.Blue dashed lines represent hydrogen bonding, and gray dashed lines represent hydrophobic interactions.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 1 .
Summary of major ABCB1/P-gp inhibitors in clinical development.

Table 3 .
MDR-reversal effects of compound 8 on ABCBl substrate drugs and non-substrate drugs.