P-glycoprotein (Pgp); multidrug resistance protein 1 (MRP-1); breast cancer resistance protein with arginine at amino acid 482 (BCRP^R482). [1]

Does not modulate BCRP^R482T (mutant with threonine at position 482). [1]

In addition to acting to reverse drug resistance induced by wild-type BCRP (BCRPR482) and to increase drug uptake and retention, biicodar has efficacy against P-glycoprotein (Pgp) and MRP-1. Bilicoda boosted mitoxantrone and daunorubicin absorption by 55% and 100%, respectively, retention rates by 100% and 60%, and cytotoxicity by 3.1-fold in 8226/Dox6 cells (Pgp). and 6.9 instances. In HL60/Adr (MRP-1) and 8226/MR20 cells (BCRP(R482)), biicodar likewise improves uptake, retention, and cytotoxicity; however, it has no effect on MCF7 AdVP3000 cells (BCRP(R482T)) [1]. A non-macrocyclic derivative of pipecolate, VX-710 interacts to the FK506 receptor protein. It has been demonstrated that VX-710 can regain sensitivity to a variety of multidrug-resistant cells, such as those found in leukemia, cancer, melanoma, and myeloma [2]. With EC50 values of 0.75 and 0.55 μM, respectively, bisicodar may efficiently block the photoaffinity labeling of P-glycoprotein by [3H]azidopine or [125I]iodoaryl azido-prazosin [3].

---

In Pgp-overexpressing 8226/Dox6 cells, VX-710 (2.5 μM) increased mitoxantrone uptake by 55% (D=0.49) and retention by 100% (D=0.53); increased daunorubicin uptake by 100% (D=0.71) and retention by 60% (D=0.53). Cytotoxicity: resistance modifying factor (RMF) of 3.1 for mitoxantrone and 6.9 for daunorubicin. [1]

In MRP-1-overexpressing HL60/Adr cells, VX-710 (2.5 μM) increased mitoxantrone uptake by 43% (D=0.71) and retention by 90% (D=0.55); increased daunorubicin uptake by 130% (D=0.82) and retention by 60% (D=0.55). RMF: 2.4 for mitoxantrone, 3.3 for daunorubicin. [1]

In BCRP^R482-overexpressing 8226/MR20 cells, VX-710 (2.5 μM) increased mitoxantrone uptake by 60% (D=0.37) and retention by 40% (D=0.34); RMF for mitoxantrone = 2.4. Daunorubicin uptake and retention increased only by 10% (D=0.17 and 0.13 respectively), yet RMF for daunorubicin was 3.6. Confocal microscopy showed no change in intracellular daunorubicin distribution (nuclear/cytoplasmic) in 8226/MR20 cells after 2 h with 5 μM daunorubicin or 24 h with 0.5 μM daunorubicin in the presence of 2.5 μM VX-710. [1]

In BCRP^R482T-overexpressing MCF7 AdVp3000 cells, VX-710 had minimal effects: mitoxantrone retention D=0.05, daunorubicin D=0.26, doxorubicin D=0.15, topotecan D=0.06; SN38 uptake (HPLC) was 2.18 ng/1×10⁶ cells with VX-710 vs. 2.9 ng without (no significant increase). RMFs: mitoxantrone 1.3, daunorubicin 1.8, doxorubicin not specified, topotecan minimal, SN38 minimal. [1]

In wild-type HL60 cells (no MDR protein overexpression), VX-710 (2.5 μM) increased mitoxantrone retention but not cytotoxicity; increased daunorubicin uptake by 20% (D=0.35) but no effect on retention (D=0.04) or cytotoxicity. [1]

Drug uptake and retention by flow cytometry: Cells (1×10⁶/ml) were incubated for 30 min at 37°C with fluorescent drug (mitoxantrone 3 μM, daunorubicin 3 μM, doxorubicin 3 μM, or topotecan 100 μM) with or without modulator (2.5 μM VX-710). For retention, after 30 min uptake without modulator, cells were resuspended in medium with or without modulator and incubated for 90 min at 37°C. Cellular fluorescence was analyzed on a flow cytometer with 488 nm excitation. Modulation was assessed by Kolmogorov-Smirnov statistic (D-value) and relative increase in mean fluorescence intensity (ΔMFI). [1]

Cytotoxicity assay (WST-1 colorimetric): Suspension cells (10,000/well) or adherent cells (600-2000/well) were plated in 96-well plates. Drugs (0.3 nM to 10 μM or up to 1 mM, half-log increments) with or without 2.5 μM VX-710 were added. After 96 h incubation, WST-1 reagent (10 μl/well) was added for 4 h, then absorbance at 450 nm (with 600 nm background) was read. IC₅₀ values and resistance modifying factor (RMF = IC₅₀ drug alone / IC₅₀ drug+modulator) were calculated. [1]

SN38 uptake by HPLC: Cells (2×10⁶) were extracted with cold acidified methanol, sonicated, centrifuged, supernatant evaporated, reconstituted, and injected into HPLC with fluorescence detection (excitation 370 nm, emission 510 nm). Mobile phase: 20% acetonitrile/80% triethylamine buffer pH 5.5. SN38 concentration measured after incubation with 2.5 μM SN38 with or without VX-710 (2.5 μM). [1]

Confocal microscopy for intracellular drug distribution: Cells (1×10⁶) were incubated with 5 μM daunorubicin for 2 h or 0.5 μM daunorubicin for 24 h, with or without 2.5 μM VX-710. After washing, cells were mounted on slides with alcian blue and evaluated by confocal microscopy (488 nm excitation, 550 nm long-pass filter). 10-20 focal planes per cell were examined. [1]

VX-710 has minimal or no effects on the pharmacokinetics of doxorubicin and mitoxantrone, but does alter paclitaxel pharmacokinetics. [1]

In clinical studies, steady-state concentrations of VX-710 (≥8 μM) were 2-3 fold higher than levels effective in vitro. [1]

VX-710 is very well tolerated. Doses producing plasma concentrations sufficient for complete MDR reversal in vitro have not been associated with toxicity. [1]

In Phase I studies, adverse reactions included headaches, mild to moderate nausea, mild reversible hypotension, and at the highest dose mild hyperbilirubinaemia. [1]

In the present study, VX-710 itself was not cytotoxic: less than 10% difference in growth was observed between cells cultured without modulator and with 2.5 μM VX-710 for 96 h in all cell lines tested. [1]

[1]. VX-710 (biricodar) increases drug retention and enhances chemosensitivity in resistant cells overexpressing P-glycoprotein, multidrug resistance protein, and breast cancer resistance protein. Clin Cancer Res. 2004 Mar 1;10(5):1826-34.

[2]. BIRICODAR (VX-710; Incel): an effective chemosensitizer in neuroblastoma. Br J Cancer. 1999 Jun;80(8):1190-6.

[3]. Cellular and biochemical characterization of VX-710 as a chemosensitizer: reversal of P-glycoprotein-mediated multidrug resistance in vitro. Anticancer Drugs. 1997 Feb;8(2):125-40.

Biricodar is an α-amino acid ester. The piperidine ester derivative Biricodar (VX-710) is a clinically used modulator of P-glycoprotein (Pgp) and multidrug resistance protein (MRP-1). Biricodar is a piperidine ester derivative. As a modulator of P-glycoprotein (P-gp) and multidrug resistance protein (MRP-1), Biricodar can restore drug sensitivity in cells expressing P-glycoprotein and MRP1. P-glycoprotein and MRP1 are efflux pumps and are the main cause of tumor cell resistance to chemotherapy drugs. This drug binds directly to Pgp and MRP-1, inhibiting efflux pump activity, thereby increasing the accumulation and retention of cytotoxic drugs within cells. Drug Indications Intravenous Biricodar citrate can be used in combination with anticancer chemotherapy drugs.

---

Mechanism of Action
Vertex's research shows that pelidazolidinyl citrate can block drug pumps P-gp and MRP, thereby enhancing the accumulation of chemotherapeutic drugs in tumor cells and restoring tumor sensitivity to chemotherapeutic drugs.

---

VX-710 (biricodar; Incel) is a pipecolinate derivative that acts as a broad-spectrum modulator of multidrug resistance (MDR) proteins. It reverses Pgp- and MRP-1-mediated resistance and also modulates wild-type BCRP (BCRP^R482) but not the mutant BCRP^R482T. [1]

VX-710 has been studied clinically in combination with paclitaxel, doxorubicin, and mitoxantrone. It has shown resensitization of paclitaxel-refractory epithelial tumors. [1]

The lack of significant pharmacokinetic interaction with anthracyclines and mitoxantrone makes it attractive for acute myeloid leukemia (AML) where anthracyclines are mainstays. [1]

C34H41N3O7

603.71

603.294

159997-94-1

3037617

Colorless to light yellow ointment

1.195g/cm3

752.1ºC at 760 mmHg

408.7ºC

1.66E-22mmHg at 25°C

1.565

4.961

0

9

16

44

866

1

O(C([C@]1([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])N1C(C(C1C([H])=C(C(=C(C=1[H])OC([H])([H])[H])OC([H])([H])[H])OC([H])([H])[H])=O)=O)=O)C([H])(C([H])([H])C([H])([H])C([H])([H])C1=C([H])N=C([H])C([H])=C1[H])C([H])([H])C([H])([H])C([H])([H])C1=C([H])N=C([H])C([H])=C1[H]

CGVWPQOFHSAKRR-NDEPHWFRSA-N

InChI=1S/C34H41N3O7/c1-41-29-20-26(21-30(42-2)32(29)43-3)31(38)33(39)37-19-5-4-16-28(37)34(40)44-27(14-6-10-24-12-8-17-35-22-24)15-7-11-25-13-9-18-36-23-25/h8-9,12-13,17-18,20-23,27-28H,4-7,10-11,14-16,19H2,1-3H3/t28-/m0/s1

1,7-Dipyridin-3-ylheptan-4-yl (2S)-1-[2-oxo-2-(3,4,5-trimethoxyphenyl)acetyl]piperidine-2-carboxylate

Biricodar Incel VX710VX-710 VX 710

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

---

Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

View More

Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

---

Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

View More

Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

---

Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

---

 (Please use freshly prepared in vivo formulations for optimal results.)