Biological Activity

Bioactivity / Mechanism

NVP-BHG712 inhibits EphB4 receptor autophosphorylation. In the original cellular study, the reported ED50 was 25 nM in EphB4-overexpressing A375 cells. This is a cellular phosphorylation endpoint; it is not a biochemical IC50 or an in vivo dose. The inhibitor also affects other Eph receptors, limiting attribution of biological effects exclusively to EphB4. [1]

Literature-reported observations and conditions; not Adooq-tested protocols or general treatment recommendations.

Target & Activity

Target	Activity	Value
EphB4	ED50	25nM
C-Raf	IC50	0.395μM
c-Src	IC50	1.266μM
c-Abl	IC50	1.667μM
Tie-2	IC50	>10μM
Axl	IC50	>10μM
HER1	IC50	>10μM
PKA	IC50	>10μM
CDK1	IC50	>10μM
PKB	IC50	>10μM
PDK-1	IC50	>10μM
FGFR3	IC50	>10μM
c-Met	IC50	>10μM

13 targets · Scroll to view more

In Vitro Studies

The original cell-based RTK assays reported ED50 values of 25 nM for EphB4 and 4200 nM for VEGFR2; IGF1R and insulin receptor ED50 values were >10,000 nM. These comparisons concern the paper’s synthesized NVP-BHG712 and do not authenticate commercial lots. [1]

Cellular EphB4 autophosphorylation inhibition

Cell-based study · A375 melanoma cells stably overexpressing human full-length EphB4; additional cellular RTK comparator assays. [1]

NVP-BHG712 inhibited cellular EphB4 phosphorylation with reported ED50 25 nM (n = 8). Cellular VEGFR2 inhibition required a higher concentration; IGF1R and insulin receptor endpoints exceeded 10,000 nM. These are cellular assay endpoints, not animal doses or biochemical IC50 values.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
EphB4 autophosphorylation in A375-EphB4 cells	ED50 25 nM (n = 8)	[1]
Cellular VEGFR2 phosphorylation	ED50 4200 nM (n = 3)	[1]
Cellular IGF1R and insulin receptor phosphorylation	ED50 IGF1R >10,000 nM (n = 3); InsR >10,000 nM (n = 3)	[1]

Literature-Reported Treatment Conditions

Treatment concentration: Concentration-response assay; numerical treatment range not given in the Figure 1 table or EphB4 ELISA method.
Exposure duration: EphrinB2-Fc stimulation for 20 min; compound exposure duration not specified in the EphB4 ELISA method.
Method: Cell-based capture ELISA for phosphorylated EphB4 after ephrinB2-Fc stimulation (1 μg/mL).
Reported limits: Compound pretreatment duration, cellular vehicle, raw dose-response curves and variance of ED50 estimates are not specified for this table. Comparator cell models/protocols are referred to prior work; conditions are not imported here.

Methods and source locations

Compound reported in the paper: NVP-BHG712

Evidence concerns the paper’s synthesized material; it does not validate the identity, purity or potency of an Adooq lot.

Figure 1 cell-based assay table; Results: Kinase inhibitor profile; Methods: Cell based phospho-RTK-ELISA; Results: NVP-BHG712 is highly potent in EphR autophosphorylation cell based assays; Figure 2 caption; Discussion: inability to discriminate among Eph receptors

EphB4 autophosphorylation in A375-EphB4 cells: Capture ELISA; ephrinB2-Fc stimulation; n is reproduced from the table without assigning an unreported replicate type. — Figure 1 cell-based assay table; Methods: Cell based phospho-RTK-ELISA

Cellular VEGFR2 phosphorylation: Cell-based phospho-RTK ELISA; detailed comparator protocol referred to previous publications. — Figure 1 cell-based assay table; Results: Kinase inhibitor profile

Cellular IGF1R and insulin receptor phosphorylation: Cellular phospho-RTK assays; no numeric estimate above the tested/reporting boundary is inferred. — Figure 1 cell-based assay table

In Vivo Studies

In a mouse VEGF-driven implant assay, oral NVP-BHG712 at 3, 10 or 30 mg/kg/day reduced tissue growth and blood content after four days. Tie-2 reductions were significant at 10 and 30 mg/kg/day. Other Eph receptors could contribute to this antiangiogenic response. [1]

Activity depended on the experimental setting. In paired A375 melanoma xenografts, synthesized NVP-BHG712 at 10 mg/kg orally once daily on weekdays did not significantly inhibit growth of mock-transfected or EphB4-overexpressing tumors. The separately synthesized NVPiso comparator likewise showed no significant growth inhibition. The two regioisomers must be distinguished when interpreting commercial-material studies. [2]

VEGF-driven angiogenesis in a mouse implant model

Animal study · Mice carrying subcutaneous VEGF-containing agar chambers. [1]

Daily oral NVP-BHG712 reduced VEGF-driven tissue growth and vascularization in a four-day implant assay. The 3 mg/kg group had significant tissue-weight and blood-content reductions; significant Tie-2 reductions were marked at 10 and 30 mg/kg, not at 3 mg/kg. No exact bar heights are extracted.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
VEGF-driven tissue weight, blood content and total Tie-2 around implanted chambers	Direction and significance 3 mg/kg/day: tissue weight and blood content reduced (p < 0.05); Tie-2 has no significance marker. 10 and 30 mg/kg/day: all three endpoints reduced (p < 0.05).	[1]

Conditions shared by the study groups

Route: Oral (p.o.)
Frequency: Once daily
Treatment course: 4-day implant assay
Vehicle: PEG-labeled control; exact treatment formulation not specified for Figure 5.

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
NVP-BHG712 dose series 3, 10 and 30 mg/kg/day	VEGF chamber + PEG control; buffer/agar chamber + PEG provides a non-VEGF baseline.	Tissue weight and blood content decreased at each tested dose with significance markers; Tie-2 decreased significantly at 10 and 30 mg/kg/day. Blood content was not monotonically lower with increasing dose.	[1]

NVP-BHG712 dose series: Model: Mice carrying subcutaneous VEGF-containing agar chambers.; Reported limits: No exact numerical bar values or group-level raw data extracted; sex/strain not assigned from another experiment.

Methods and source locations

Compound reported in the paper: NVP-BHG712

Evidence concerns the paper’s synthesized material; it does not validate the identity, purity or potency of an Adooq lot.

Figure 5a–c and caption; Results: Inhibition of EphB4 forward signaling inhibits VEGF driven angiogenesis; Methods: In vivo growth factor-induced angiogenesis model

VEGF-driven tissue weight, blood content and total Tie-2 around implanted chambers: Mean ± SEM; n = 6 per group per experiment; Mann–Whitney Rank Sum Test, as stated in the caption. Numeric effect sizes not tabulated and not estimated from bars. — Figure 5a–c image and caption

NVP-BHG712 dose series · Figure 5a–c and caption

NVP versus NVPiso: no significant A375 xenograft-growth inhibition

Animal study · Rj:NMRI-Foxn1 nu/nu mice bearing paired A375-pIRES and EphB4-overexpressing A375 tumors. [2]

At 10 mg/kg orally on weekdays, neither synthesized NVP nor NVPiso significantly inhibited growth of either A375 tumor type versus vehicle. A slight increase in growth was described as a trend. EphB4-overexpressing tumors grew faster than mock-transfected tumors; that within-mouse comparison is distinct from a treatment effect.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
Growth of A375-pIRES and A375-EphB4 xenografts versus vehicle	Reported treatment effect Neither NVP nor NVPiso significantly affected growth; slight increase described as a trend. Figure 2a,b: mean ± SEM, n = 14–15.	[2]

Conditions shared by the study groups

Route: Oral
Frequency: Once a day, weekdays only
Treatment course: Day 1 after tumor-cell injection until experiment end; growth curves through day 13.
Vehicle: 10% v/v 1-methyl-2-pyrrolidone and 90% v/v PEG300; 6.7 mL/kg body weight.

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
NVP 10 mg/kg body weight	Same-volume vehicle-treated mice; each mouse carries mock and EphB4-overexpressing tumors.	No significant inhibition of either tumor type versus vehicle; slightly increased growth described as a trend.	[2]
NVPiso — distinct regioisomer comparator 10 mg/kg body weight	Same-volume vehicle-treated mice; each mouse carries mock and EphB4-overexpressing tumors.	No significant inhibition of either tumor type versus vehicle; slightly increased growth described as a trend.	[2]

NVP: Model: Rj:NMRI-Foxn1 nu/nu mice bearing paired A375-pIRES and EphB4-overexpressing A375 tumors.; Reported limits: No exact between-treatment p value or tumor exposure/target-engagement measurement provided for this endpoint.

NVPiso — distinct regioisomer comparator: Model: Rj:NMRI-Foxn1 nu/nu mice bearing paired A375-pIRES and EphB4-overexpressing A375 tumors.; Reported limits: No exact between-treatment p value or tumor exposure/target-engagement measurement provided for this endpoint.

Methods and source locations

Compound reported in the paper: NVP-BHG712; Compound label in the paper: NVP

Apply NVP results to the named original structure only. NVPiso comparator findings do not establish activity of NVP-BHG712 or any Adooq lot.

Results 2.1; Figure 2a–c and caption; Methods 4.1, 4.3, 4.4, 4.9

Growth of A375-pIRES and A375-EphB4 xenografts versus vehicle: Caliper measurements; V = π/6 × (length × width²). No curve-derived exact tumor volumes or unreported treatment p values inferred. — Results 2.1; Figure 2a,b and caption

NVP · Results 2.1; Figure 2a,b; Methods 4.1, 4.3, 4.4

NVPiso — distinct regioisomer comparator · Results 2.1; Figure 2a,b; Methods 4.1, 4.3, 4.4

Specifications

Manufacturer	Adooq Bioscience
Actions	Inhibitor
CAS Number	940310-85-0
Applications	EphB4 inhibitor
Formula	C₂₆H₂₀F₃N₇O
Molecular Weight	503.5
Synonyms	BHG712
SMILES	CC1=C(C=C(C=C1)C(=O)NC2=CC=CC(=C2)C(F)(F)F)NC3=NC(=NC4=C3C=NN4C)C5=CN=CC=C5

Storage & Solubility

In Vitro Solubility

Reported Solubility

Solvent	Reported Solubility
DMSO	90 mg/mL (178.75 mM)
Water	Insoluble
Ethanol	3 mg/mL (5.95 mM)

Preparing Stock Solutions

Stock preparation is unavailable for this product molecular weight.

Calculated stock concentrations do not establish solvent solubility. Refer to the reported solubility data for experimental guidance.

In Vivo Preparation

Concentration: 28 mg/mL
Vehicle: NMP+PEG300 (10+90, v+v)

Storage

Store powder at -20°C, keep desiccated.
In powder form, the chemical is stable for 36 months.
In solution, store at -20°C and use within 1 month to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.

Research References

[1] Georg Martiny-Baron, Philipp Holzer, Eric Billy, et al. The small molecule specific EphB4 kinase inhibitor NVP-BHG712 inhibits VEGF driven angiogenesis Angiogenesis (2010). PMID: 20803239 DOI: 10.1007/s10456-010-9183-z Source material
[2] Christin Neuber, Alix Tröster, Reik Löser, et al. The Pyrazolo[3,4-d]pyrimidine-Based Kinase Inhibitor NVP-BHG712: Effects of Regioisomers on Tumor Growth, Perfusion, and Hypoxia in EphB4-Positive A375 Melanoma Xenografts Molecules (Basel, Switzerland) (2020). PMID: 33153234 DOI: 10.3390/molecules25215115 Source material

Publications

2 publications citing this product

Ryuzaburo Yuki, .et al. , Eur J Pharmacol, 2024, Jan 15:963:176229 PMID: 38072041
Y Kaibori, .et al. , the FASEB Journal, 2019, Jan 22:fj201801519RR PMID: 30668924

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NVP-BHG712

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NVP-BHG712 1 mg

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Biological Activity

Bioactivity / Mechanism

Target & Activity

In Vitro Studies

Cellular EphB4 autophosphorylation inhibition

Literature-Reported Treatment Conditions

In Vivo Studies

VEGF-driven angiogenesis in a mouse implant model

Conditions shared by the study groups

NVP versus NVPiso: no significant A375 xenograft-growth inhibition

Conditions shared by the study groups

Specifications

Storage & Solubility

In Vitro Solubility

Reported Solubility

Preparing Stock Solutions

In Vivo Preparation

Storage

Research References

Publications

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