Lysine-specific demethylase 1 (LSD1)
Bomedemstat is a potent and irreversible inhibitor of lysine-specific demethylase 1 (LSD1), an enzyme that removes methyl groups from histone H3 at lysine 4 (H3K4) and lysine 9 (H3K9). By inhibiting LSD1, bomedemstat increases H3K4 and H3K9 methylation levels, thereby altering the expression of genes involved in cell proliferation, differentiation, and survival. LSD1 inhibition leads to cell cycle arrest at the S phase and induces apoptosis in cancer cells. Bomedemstat is an orally active compound with antineoplastic activity.

Bomedemstat increases p53 expression and methylation while concurrently inhibiting proliferation and inducing apoptosis in Jak2V617F cells[1]. Cell cycle arrest and enhanced survival are caused by bomedemstat (50 nM-1 μM; 96 h), which also promotes apoptosis via BCL-XL and PUMA in a TP53-dependent manner[1].
Bomedemstat induces cell cycle arrest at the S phase in SET-2 acute megakaryoblastic leukemia cells and induces apoptosis in Ba/F3 murine hematopoietic cells expressing Jak2V617F when used at a concentration of 1 μM. The compound increases H3K4 and H3K9 methylation and alters gene expression. Bomedemstat inhibits cancer cell proliferation and induces apoptosis. In SCLC syngeneic models, bomedemstat potentiates the response to PD-1 inhibition, suggesting potential for combination with immunotherapy.

Treatment with bomedemstat (oral gavage; 45 mg/kg; once daily; 56 d) improves or returns blood cell counts to normal, decreases spleen sizes, reestablishes normal splenic architecture, and lessens fibrosis in the bone marrow[1].
In vivo, bomedemstat demonstrates antitumor activity in various preclinical models. In syngeneic mouse models of small cell lung cancer (SCLC), bomedemstat potentiates the response to PD-1 inhibition, leading to enhanced antitumor efficacy. The compound inhibits cancer cell proliferation and induces apoptosis in vivo. Bomedemstat is being investigated for the treatment of myelodysplastic syndrome, myelofibrosis, and other hematologic malignancies. Its oral bioavailability supports convenient dosing in preclinical and clinical studies.

LSD1 enzymatic activity assays are performed using recombinant LSD1 enzyme and histone H3 peptide substrates (typically residues 1-21) with methylation at lysine 4. The demethylase activity is measured by detecting the production of formaldehyde or by using antibody-based detection of demethylated product. Bomedemstat is incubated with the enzyme and substrate in assay buffer (50 mM HEPES, pH 7.5, containing 0.01% Triton X-100 and 2 mM DTT) for 30-60 minutes at 37°C. The reaction is stopped and product formation is quantified. IC50 values are calculated from concentration-response curves. The irreversible nature of inhibition is confirmed by dilution or dialysis experiments.

Apoptosis Analysis[1]
Cell Types: SET-2 cells
Tested Concentrations: 50 nM, 100 nM, and 1 μM
Incubation Duration: 96 hrs (hours)
Experimental Results: diminished levels of the antiapoptotic protein BCL-XL and increased levels of the pro-apoptotic protein PUMA.
Cellular assays for LSD1 inhibition typically employ cancer cell lines sensitive to LSD1 inhibition, such as acute megakaryoblastic leukemia (SET-2) cells or small cell lung cancer cells. Cells are treated with varying concentrations of bomedemstat for 24-72 hours. Cell viability is assessed using MTT or CellTiter-Glo assays. Apoptosis is evaluated by Annexin V/PI staining and caspase activity assays. Cell cycle analysis is performed by propidium iodide staining and flow cytometry. Histone methylation changes (H3K4me2, H3K9me2) are assessed by Western blot. Gene expression changes are analyzed by qPCR or RNA-seq.

Animal/Disease Models: Mx-Jak2V617F mice[1]
Doses: 45 mg/kg
Route of Administration: po (oral gavage); 45 mg/kg; one time/day; 56 days
Experimental Results: decreased splenomegaly Dramatically with a few treated mice normalizing their spleen weight, the 56-day course led to partial restoration of lymph follicles and spleen architecture by histological examination.
In vivo efficacy is evaluated in murine xenograft or syngeneic tumor models. Tumor-bearing mice are treated with bomedemstat orally at various doses (typically 10-50 mg/kg) daily. Tumor volumes are measured every 2-3 days. Combination studies with PD-1/PD-L1 inhibitors are conducted to assess synergistic efficacy. Pharmacodynamic markers such as H3K4 and H3K9 methylation levels are measured in tumor tissues to confirm target engagement. Body weight and clinical signs are monitored for tolerability assessment.

Bomedemstat is orally active with good bioavailability. The compound is available as the ditosylate salt for improved physicochemical properties. Standard pharmacokinetic studies in preclinical species would characterize absorption, distribution, metabolism, and excretion parameters. The compound's oral bioavailability supports convenient dosing in clinical studies. Bomedemstat is being investigated in clinical trials for hematologic malignancies and solid tumors.

Preclinical toxicology studies for bomedemstat are ongoing as part of clinical development. As an irreversible LSD1 inhibitor, potential toxicities may include myelosuppression, gastrointestinal effects, and other on-target effects related to altered gene expression. The compound has been evaluated in preclinical models with acceptable tolerability. Standard safety pharmacology, genotoxicity, and repeated-dose toxicity studies would be required for regulatory approval.

[1]. LSD1 Inhibition Prolongs Survival in Mouse Models of MPN by Selectively Targeting the Disease Clone. Hemasphere.2018 Jun 8;2(3):e54.

[2]. LSD1/KDM1A inhibitors in clinical trials: advances and prospects. J Hematol Oncol. 2019 Dec 4;12(1):129.

IMG-7289 is currently undergoing clinical trial NCT03136185 (IMG-7289 for the treatment of patients with myelofibrosis).
Bromidesistat is an oral, irreversible lysine-specific demethylase 1 (LSD1) inhibitor with potential antitumor activity. After administration, bromedistat binds to LSD1 and inhibits its activity. LSD1 is a demethylase that inhibits the expression of target genes by converting the dimethylated and monomethylated forms of histone H3 lysine 4 (H3K4) to monomethylated and unmethylated H3K4. LSD1 inhibition enhances H3K4 methylation and increases the expression of tumor suppressor genes. Furthermore, LSD1 can demethylate monomethylated or dimethylated H3K9, thereby increasing the expression of pro-tumorigenic genes; therefore, inhibiting LSD1 can also promote H3K9 methylation and reduce the transcription of these genes. In summary, this may lead to the inhibition of growth in LSD1-overexpressing tumor cells. LSD1 is an enzyme belonging to the flavin adenine dinucleotide (FAD)-dependent amine oxidase family, overexpressed in certain tumor cells, and plays a crucial role in the regulation of gene expression, tumor cell growth, and survival. Despite recent advances, myeloproliferative neoplasms (MPNs) still have a considerably high morbidity and mortality rate. Janus kinase (JAK) inhibitors, such as ruxotinib, can control symptoms but do not significantly alter the natural course of the disease. In this report, we demonstrate the effect of IMG-7289 (an irreversibly epigenetically active lysine-specific demethylase 1 (LSD1) inhibitor) in a mouse model of MPN. Once-daily treatment with IMG-7289 normalized or improved blood cell counts, reduced spleen volume, restored normal spleen structure, and alleviated myelofibrosis. More importantly, the LSD1 inhibitor reduced the mutant allele load and improved survival. IMG-7289 selectively inhibits the proliferation of JAK2 V617F cells and induces their apoptosis by simultaneously increasing the expression and methylation of p53, as well as independently increasing the expression and methylation of the pro-apoptotic factor PUMA, and decreasing the level of its anti-apoptotic antagonist BCLXL. These data provide a molecular understanding of the disease-modifying activity of the LSD1 inhibitor IMG-7289, which is currently being clinically evaluated in high-risk myelofibrosis patients. In addition, low-dose IMG-7289 and ruxolitinib have a synergistic effect in mice, normalizing the MPN phenotype, which provides a theoretical basis for studying combination therapy. [1]

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Histone demethylase LSD1 plays a key role in tumorigenesis, and targeting LSD1 is becoming an emerging option for cancer treatment. To date, a number of LSD1 inhibitors have been reported, some of which, such as TCP, ORY-1001, GSK-2879552, IMG-7289, INCB059872, CC-90011 and ORY-2001, are currently undergoing clinical evaluation for cancer treatment, particularly for small cell lung cancer (SCLC) and acute myeloid leukemia (AML). This review aims to provide a comprehensive overview of clinical trials of LSD1 inhibitors, including studies on molecular mechanisms, clinical efficacy, adverse reactions and pharmacodynamic/pharmacokinetic studies, and to provide an outlook on the future development of this field. [2]
Bomedemstat (IMG-7289) is being developed for the treatment of myelodysplastic syndrome, myelofibrosis, and other hematologic malignancies. It has also shown activity in small cell lung cancer models, particularly in combination with PD-1 inhibitors. The compound is a potent and irreversible LSD1 inhibitor that increases H3K4 and H3K9 methylation. Clinical trials are ongoing to evaluate its safety and efficacy in various indications.

C42H50FN7O8S2

864.016911029816

863.314

C, 58.39; H, 5.83; F, 2.20; N, 11.35; O, 14.81; S, 7.42

1990504-72-7

Bomedemstat hydrochloride;Bomedemstat dihydrochloride;Bomedemstat;1990504-34-1; 1990504-72-7

131801105

White to off-white solid powder

4

13

12

60

986

3

CC1=CC=C(C=C1)S(=O)(=O)O.CC1=CC=C(C=C1)S(=O)(=O)O.CN1CCN(CC1)C(=O)[C@H](CCCN[C@@H]2C[C@H]2C3=CC=C(C=C3)F)NC(=O)C4=CC=C(C=C4)N5C=CN=N5

WLRWBXFBIJMKHG-DVNXTAPYSA-N

InChI=1S/C28H34FN7O2.2C7H8O3S/c1-34-15-17-35(18-16-34)28(38)25(3-2-12-30-26-19-24(26)20-4-8-22(29)9-5-20)32-27(37)21-6-10-23(11-7-21)36-14-13-31-33-36;2*1-6-2-4-7(5-3-6)11(8,9)10/h4-11,13-14,24-26,30H,2-3,12,15-19H2,1H3,(H,32,37);2*2-5H,1H3,(H,8,9,10)/t24-,25-,26+;;/m0../s1

N-[(2S)-5-[[(1R,2S)-2-(4-fluorophenyl)cyclopropyl]amino]-1-(4-methylpiperazin-1-yl)-1-oxopentan-2-yl]-4-(triazol-1-yl)benzamide;4-methylbenzenesulfonic acid

IMG7289 ditosylate; IMG-7289; IMG 7289; BOMEDEMSTAT DITOSYLATE; IMG-7289 ditosylate; 1990504-72-7; Bomedemstat tosylate; Bomedemstat (ditosylate); Bomedemstat ditosylate (IMG 7289); 2PYF2U9JNF; Bomedemstat tosylate [USAN]; Bomedemstat ditosylate

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

DMSO : ~100 mg/mL (~115.74 mM)

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.

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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).

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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)

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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).

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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

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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.

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