CDK2/cyclinE (IC50 = 4.6 μM); cdk2/cyclin A (IC50 = 83 μM); CDK9/cyclinT1 (IC50 = 2.7 μM); ERK2 (IC50 = 52 μM)
Bohemine is a synthetic cyclin-dependent kinase (CDK) inhibitor. It is a 2,6,9-trisubstituted purine derivative and an analog of aromatic cytokinins. The mechanism of action involves competition with ATP for binding to CDKs. The study indicates it perturbs cell cycle progression by acting at both the G1/S and G2/M boundaries, but specific IC50, Ki, or EC50 values for individual CDKs are not provided in this literature. [1]

Bohemine treatment inhibits cell growth. When bohemine is added at concentrations between 1 and 10 µM, growth and the production of monoclonal antibodies are temporarily stopped. Following the brief suppression of cell functions, there is a notable, transient increase in both the rate of specific growth and production[1].
The G1/S and G2/M boundaries of hybridoma cells are retarded, based on the concentration of bohemine (0-30 µM)[1].
T-cell lymphoblastic line Five proteins are shown to be downregulated when bohemine is used to treat CEM: α-enolase, triosephosphate isomerase, initiation factor 5A, and the α- and β-subunits of Rho GDP-dissociation inhibitor 1. These proteins are important for the processes of glycolysis, proteosynthesis, and cytoskeleton rearrangement[1].
With an IC50 of 27 µM, bohemine inhibits the growth of human tumor cell lines[2].

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In pilot studies using mouse hybridoma ME-750 cell cultures, bohemine at concentrations up to 30 µM did not induce apoptosis, which distinguished it from many other antiproliferative agents. [1]
Addition of bohemine (1-10 µM) to batch cultures of hybridoma cells resulted in an initial, short-term arrest of cell growth and monoclonal antibody (MAb) production within 24 hours. This suppression phase was followed by a significant temporary increase in both the specific growth rate and specific MAb production rate, which in most cases exceeded the rates in untreated control cultures. [1]
In static and fed-batch cultures initiated at high cell density, bohemine at 10 and 30 µM significantly suppressed cell growth. At lower concentrations (1 and 3 µM), viable cell densities after day 5 were slightly higher than in controls, especially in fed-batch mode. Final MAb concentrations in fed-batch cultures with 3 µM bohemine showed an approximately 18% increase relative to control, though this was not statistically significant (P < 0.05). Bohemine at 30 µM inhibited both growth and MAb synthesis. [1]
In semicontinuous cultures (chemostat-like mode), steady-state viable cell density showed a slight stimulatory effect at micromolar concentrations of bohemine, but significant inhibition at 10 and 30 µM. The same trend was observed for steady-state MAb concentrations. [1]
Cell cycle analysis in semicontinuous cultures revealed two concentration-dependent effects. The major trend was an increase in the fraction of cells in the G0/G1 phase at the expense of the S phase fraction, indicating a block at the G1/S transition. A minor trend was an increase in G2/M phase cells at 1 and 3 µM bohemine, indicating retardation at the G2/M transition. The level of apoptotic cells remained relatively low and constant across all concentrations tested. [1]

Bohemine (50 mg/kg; intravenous injection; BALB/c mice) treatment reveals a T1/2 of 1.39 hours, an observed clearance of 0.23 L/h, and a Cmax of 72,308 nM[2].

The study employed mouse hybridoma cell line ME-750. Cells were cultured in a DMEM/F12/RPMI 1640 (2:1:1) based medium supplemented with amino acids, glutamine, HEPES, sodium bicarbonate, and a protein-free growth-promoting mixture. [1]
Experiments were conducted in various culture modes: static T-flask cultures, stirred spinner flask batch cultures, fed-batch cultures (with daily nutrient feeding), and semicontinuous cultures (where a fixed volume of culture was replaced daily with fresh medium containing the test substance to maintain a constant concentration). Cultures were maintained at 37°C in a humidified atmosphere of 5% CO2. [1]
Viable and dead cell counts were determined daily using the trypan blue exclusion method with a hemocytometer. Monoclonal antibody concentration in the culture supernatant was quantified by immunoturbidimetry. [1]
For cell cycle analysis, cells were permeabilized and stained with propidium iodide using a commercial DNA staining kit. The DNA content was then measured by flow cytometry. Data were analyzed using specialized software to determine the percentage of cells in G0/G1, S, and G2/M phases, as well as the subdiploid (apoptotic) fraction. [1]

BALB/c mice bearing the colon 26 murine tumor[2]
50 mg/kg
Intravenous injection (Pharmacokinetic Analysis)

[1]. Diverse effects of the cyclin-dependent kinase inhibitor bohemine: Concentration- and time-dependent suppression or stimulation of hybridoma culture. Cytotechnology. 2001 Jul;36(1-3):117-23.

[2]. In vitro and in vivo pharmacokinetic-pharmacodynamic relationships for the trisubstituted aminopurine cyclin-dependent kinase inhibitors olomoucine, bohemine and CYC202. Clin Cancer Res. 2005 Jul 1;11(13):4875-87.

Bohemine is a purine derivative with (3-hydroxypropyl)amino, benzylamino, and isopropyl groups substituted at C-2, C-6, and N-9, respectively; it is a synthetic, cell-membrane-transmissible cyclin-dependent kinase (CDK) inhibitor with a structure similar to olomoucine and roscovitine. It is an EC 2.7.11.22 (cyclin-dependent kinase) inhibitor. Bohemine (2-[(3-hydroxypropyl)amino]-6-benzylamino-9-isopropylpurine) is an analog of the plant hormone cytokinin and a member of a library of 2,6,9-trisubstituted purine derivatives, which are known CDK inhibitors. [1] It has been preclinically studied as a potential anticancer drug. Its effects are considered pleiotropic. Proteomic analysis of T lymphoblast cells treated with bromodipine showed downregulation of multiple proteins involved in glycolysis, protein synthesis, and cytoskeleton remodeling, indicating that its cellular effects are not limited to simple CDK inhibition. [1]
This study highlights a unique biphasic response in hybridoma cells: initial function is suppressed, followed by spontaneous recovery, and growth and productivity are stimulated even without removal of the drug from the culture medium. [1]

C18H24N6O

340.42276

340.201

C, 63.51; H, 7.11; N, 24.69; O, 4.70

189232-42-6

189232-42-6

2422

White to off-white solid powder

1.3±0.1 g/cm3

589.4±60.0 °C at 760 mmHg

310.3±32.9 °C

0.0±1.7 mmHg at 25°C

1.653

1.02

3

6

8

25

390

0

CC(C)N1C=NC2=C(NCC3=CC=CC=C3)NC(=NCCCO)N=C21

OPQGFIAVPSXOBO-UHFFFAOYSA-N

InChI=1S/C18H24N6O/c1-13(2)24-12-21-15-16(20-11-14-7-4-3-5-8-14)22-18(23-17(15)24)19-9-6-10-25/h3-5,7-8,12-13,25H,6,9-11H2,1-2H3,(H2,19,20,22,23)

3-[[6-(benzylamino)-9-propan-2-ylpurin-2-yl]amino]propan-1-ol

Bohemine

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)

DMSO: 68~100 mg/mL (199.8~293.8 mM)
Ethanol: ~68 mg/mL (199.8 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (7.34 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (7.34 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
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.

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Solubility in Formulation 3: ≥ 2.5 mg/mL (7.34 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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