Protein arginine methyltransferases; PRMT1/6/8
PRMT1 (protein arginine methyltransferase 1), PRMT6 (protein arginine methyltransferase 6), and PRMT8 (protein arginine methyltransferase 8). DCPT1061 exhibits less inhibitory effect on PRMT3, PRMT4 (also known as CARM1), and PRMT5, as well as other epigenetic enzymes. PRMT1 is the major type I PRMT responsible for asymmetric dimethylation of arginine residues on histone H4 (H4R3me2a) and various non-histone proteins, regulating gene expression and protein function. PRMT6 and PRMT8 are also type I PRMTs with distinct substrate specificities and tissue expression patterns. The selective inhibition of these three isoforms, while sparing PRMT3 (which regulates ribosomal processing), PRMT4/CARM1 (which methylates histone H3), and PRMT5 (a type II PRMT that generates symmetric dimethylation), allows researchers to dissect the specific contributions of type I PRMTs to oncogenic signaling without confounding effects from other PRMT family members.

In this study, researchers found that PRMT1 expression was remarkably upregulated in tumor tissues and associated with poor pathologic characters and outcomes of ccRCC patients. Furthermore, genetic knockdown and pharmacological inhibition of PRMT1 by a novel potent inhibitor DCPT1061 dramatically induced G1 cell cycle arrest and suppressed ccRCC cell growth. Mechanistically, RNA sequencing and further validation identified Lipocalin2 (LCN2), a secreted glycoprotein implicated in tumorigenesis, as a crucial regulator of ccRCC growth and functional downstream effector of PRMT1. Epigenetic silencing of LCN2 autocrine secretion by PRMT1 deficiency decreased downstream p-AKT, leading to reduced p-RB and cell growth arrest through the neutrophil gelatinase associated lipocalin receptor (NGALR). [2]

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In this study, researchers found that protein arginine methyltransferase 1 (PRMT1) expression was inversely correlated with the number and activity of CD8+ T cells within melanoma specimen[1].

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DCPT1061 potently inhibits PRMT1, PRMT6, and PRMT8 in vitro with less inhibitory effect on PRMT3, PRMT4, and PRMT5 or other epigenetic enzymes. While the exact IC50 values for each isoform are not published in the available literature, the compound demonstrates significant biological activity with a selectivity profile that distinguishes it from pan-PRMT inhibitors. In enzymatic assays, DCPT1061 reduces the methyltransferase activity of its target PRMTs, thereby decreasing the levels of asymmetric arginine dimethylation on histone and non-histone substrates. This activity is expected to modulate downstream gene expression programs and signaling pathways that are driven by PRMT-mediated methylation, contributing to its antitumor effects. The compound's selectivity for PRMT1, PRMT6, and PRMT8 over other PRMT isoforms and epigenetic enzymes suggests a favorable profile for mechanistic studies of type I PRMT biology.

PRMT1 deficiency or inhibition with DCPT1061 significantly restrained refractory melanoma growth and increased intratumoral CD8+ T cells in vivo. Moreover, PRMT1 deletion in melanoma cells facilitated formation of double-stranded RNA derived from endogenous retroviral elements (ERV) and stimulated an intracellular interferon response. Mechanistically, PRMT1 deficiency repressed the expression of DNA methyltransferase 1 (DNMT1) by attenuating modification of H4R3me2a and H3K27ac at enhancer regions of Dnmt1, and DNMT1 downregulation consequently activated ERV transcription and the interferon signaling. Importantly, PRMT1 inhibition with DCPT1061 synergized with PD-1 blockade to suppress tumor progression and increase the proportion of CD8+ T cells as well as IFNγ+CD8+ T cells in vivo. [1]

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Moreover, PRMT1 inhibition by DCPT1061 not only inhibited tumor growth but also sensitized ccRCC to sunitinib treatment in vivo by attenuating sunitinib-induced upregulation of LCN2-AKT-RB signaling. Conclusion: Taken together, our study revealed a PRMT1-dependent epigenetic mechanism in the control of ccRCC tumor growth and drug resistance, indicating PRMT1 may serve as a promising target for therapeutic intervention in ccRCC patients[2].

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DCPT1061 has antitumor effects, demonstrating potential for cancer therapy through modulation of histone methylation pathways. In preclinical models, inhibition of PRMT1 by DCPT1061 is expected to reduce the proliferation and survival of cancer cells that depend on PRMT1 activity for oncogenic signaling. PRMT1 has been shown to regulate key pathways including the NF-κB, Wnt/β-catenin, and STAT3 signaling cascades, which are frequently dysregulated in cancer. By inhibiting PRMT1, DCPT1061 may suppress these pathways and induce apoptosis or cell cycle arrest in tumor cells. The additional inhibition of PRMT6 and PRMT8, which have also been implicated in cancer progression, may provide complementary antitumor effects. However, detailed in vivo efficacy data (such as tumor growth inhibition in xenograft models) are not provided in the available literature, and further studies are needed to fully characterize the compound's antitumor potential in animal models.

In vitro enzyme inhibition assays are performed using recombinant PRMT enzymes (PRMT1, PRMT6, PRMT8, and other isoforms for selectivity profiling) incubated with varying concentrations of DCPT1061 and a methyl donor substrate, typically S-adenosylmethionine (SAM) or radiolabeled 3H-SAM, along with a suitable peptide or protein substrate that can be methylated by the target PRMT. The reaction is carried out under optimized buffer conditions (e.g., Tris-HCl pH 8.0, NaCl, DTT) at 30°C for a specified incubation time. After the reaction is quenched (e.g., by addition of cold trichloroacetic acid or by spotting onto filter paper), the transferred methyl groups are measured via scintillation counting, filter-binding methods, or fluorescence-based detection (e.g., using methyltransferase activity kits). IC50 values for each PRMT isoform are calculated by fitting dose-response curves to the inhibition data. Selectivity is assessed by comparing the IC50 values across PRMT1, PRMT6, PRMT8, PRMT3, PRMT4, PRMT5, and other epigenetic enzymes.

The cultured cells were treated with DCPT1061 or shPRMT1 lentivirus respectively in vitro in 10cm dish plates. 48 h later, cells were collected, and total RNA was isolated. The well-constructed cDNA libraries were then sequenced on the Illumina HiSeq2000 using paired-end methods. After careful quality control of raw reads including trimming adapter sequences and filtering raw reads with low quality, the clean reads were then analyzed through a routine RNA sequencing analysis pipeline. The sequencing reads were first aligned to human hg19 genome by STAR 2.5 (Spliced Transcripts Alignment to a Reference) and then featureCounts software was used to quantify gene expression. Based on these raw counts, differential gene expression analysis was conducted using R/Bioconductor package DESeq2. To define genes differentially expressed in each sample, both fold change of 2 and an adjusted P-value of 0.05 were set as the cut-off value in each case[2].

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Cancer cell lines (e.g., various tumor types depending on the research focus) are treated with DCPT1061 at graded concentrations (typically ranging from nanomolar to micromolar) for 72-96 hours to assess antiproliferative effects. Cell viability is measured using standard colorimetric assays such as MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) or CCK-8 (Cell Counting Kit-8), which measure the metabolic activity of viable cells. The absorbance is read at 540-570 nm (MTT) or 450 nm (CCK-8), and IC50 values are calculated from dose-response curves. Additional cellular assays may include assessment of apoptosis (Annexin V/PI staining, caspase activation), cell cycle analysis (flow cytometry), and measurement of PRMT substrate methylation levels (e.g., histone H4R3me2a) by Western blotting to confirm target engagement at the cellular level.

Mouse tumor model[1]
Female 6–8-week-old BALB/c nude mice and female 6- to 8-week-old C57BL/6J wild-type (WT) mice were used. For genetic intervention, 2.5×105 B16-F10 cells for PRMT1 sg1 and CTRL were subcutaneously administrated into WT mice or nude mice. The anti-CD8α antibody (200 μg, intraperitoneal injection) were delivered to mice bearing CTRL or PRMT1 sg1 on day 0, 4, 8, 12. Tumor size and mice body weight were measured every 3 days. When tumors reached 1,000 to 1,500 mm3, mice were sacrificed. For pharmacological intervention, 2.5×105 B16-F10 cells were subcutaneously injected into WT mice or nude mice. Once the tumor volume had reached 50 to 100 mm3, the mice were randomly grouped and intraperitoneal administrated with either vehicle (PBS) or 30 mg/kg DCPT1061 once daily. The anti–PD-1 antibody (150 μg, intraperitoneal injection) were delivered to mice on day 8, 12 alone or partnered with DCPT1061. Tumor size and mice body weight were measured every 3 days. When tumors reached 1,000 to 1,500 mm3, mice were sacrificed. For survival analysis, the mice were considered dead when tumor volume reach to 2,000 mm3. A formula was used to calculate the tumor volume: 1/2 × length × width2.

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Xenograft model and treatments[2]
Around 5 × 106 ccRCC cells were injected subcutaneously into the flank region of six-week-old female nude mice (BALB/c-nu/nu). The PDX was established in six-week-old female SCID mice using the PDX#1002523691 tissue. Tumor volume was measured with a caliper, and the estimated tumor volume = length × width2 /2. When tumors reached approximately 50 mm3, four groups (n = 6) were divided randomly. Treatment of mice was as follows: vehicle control, DCPT1061 (30 mg/kg/day), sunitinib (25 mg/kg/day), and a combination of DCPT1061 (30 mg/kg/day) and sunitinib (25 mg/kg/day). Body weights and tumor volumes were measured every second day. After treatment, mice were sacrificed and the tumors were harvested, weighed, fixed with 4% formaldehyde.

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Mice bearing xenograft tumors (e.g., from human cancer cell lines) are administered DCPT1061 via intraperitoneal or oral routes at various dosages (typically multiple dose levels to establish dose-response relationships). Tumor volume is measured regularly using calipers (length × width²/2), and body weight is monitored to assess tolerability. Treatment is typically continued for 2-4 weeks or until tumors reach a predetermined size. At study endpoint, tumors are excised for histopathological analysis (e.g., H&E staining) and biomarker evaluation (e.g., immunohistochemistry for PRMT substrate methylation, Ki-67 for proliferation, cleaved caspase-3 for apoptosis) to assess in vivo target engagement and antitumor efficacy. Pharmacodynamic markers are measured to confirm that the compound reaches the tumor tissue and inhibits its targets at the achieved doses.

Predicted to have moderate oral bioavailability and reasonable plasma protein binding typical of small-molecule PRMT inhibitors. Metabolic stability is likely mediated by CYP450 enzymes in the liver, primarily through oxidative metabolism. Standard pharmacokinetic parameters (Cmax, Tmax, AUC, t1/2, clearance, volume of distribution) are typically evaluated in rodent models (mice or rats) following both intravenous and oral administration at relevant dose levels. The compound is typically formulated in suitable vehicles (e.g., DMSO/PEG400/saline mixtures) for parenteral administration or in appropriate oral formulations for gavage studies. Detailed PK data specific to DCPT1061 are not available in the published literature and would need to be generated experimentally.

Predicted to have an acceptable safety profile in preclinical species at therapeutic doses, with potential dose-limiting toxicities related to on-target PRMT inhibition affecting normal cellular methylation processes. PRMT1 is essential for embryonic development and plays important roles in normal cellular functions, so chronic inhibition may affect normal tissues, particularly those with high rates of cell turnover (e.g., bone marrow, intestinal epithelium). Standard toxicology studies (acute and sub-chronic) in rodents and non-rodents are typically required to establish the safety margin, identify target organs of toxicity, and determine the maximum tolerated dose (MTD) before advancing to clinical development. As a research compound, detailed toxicology data are not publicly available.

[1].PRMT1 is a novel molecular therapeutic target for clear cell renal cell carcinoma. Theranostics. 2021 Mar 12;11(11):5387-5403.

[2].PRMT1 inhibition activates the interferon pathway to potentiate antitumor immunity and enhance checkpoint blockade efficacy in melanoma. Cancer Res. 2024 Feb 1;84(3):419-433.

Despite the significant success of immune checkpoint blockade (ICB) in cancer treatment, many tumors, including melanoma, still exhibit innate or adaptive resistance. Tumor-inherent T-cell defects and T-cell dysfunction have been identified as key factors contributing to ICB resistance. This study found that the expression of protein arginine methyltransferase 1 (PRMT1) was negatively correlated with the number and activity of CD8+ T cells in melanoma specimens. PRMT1 deficiency or inhibition using DCPT1061 significantly suppressed the growth of refractory melanoma and increased the number of intratumoral CD8+ T cells in vivo. Furthermore, PRMT1 deficiency in melanoma cells promoted the formation of endogenous retroviral element (ERV)-derived double-stranded RNA and stimulated intracellular interferon responses. Mechanistically, PRMT1 deficiency inhibits the expression of DNA methyltransferase 1 (DNMT1) by attenuating the H4R3me2a and H3K27ac modifications of the DNMT1 enhancer region, and the downregulation of DNMT1 subsequently activates the transcriptional and interferon signaling pathways of endogenous retroviruses (ERVs). Importantly, the synergistic effect of PRMT1 inhibition by DCPT1061 with PD-1 blockers inhibits tumor progression and increases the proportion of CD8+ T cells and IFNγ+CD8+ T cells in vivo. In summary, these results reveal an unknown role and mechanism of PRMT1 in regulating anti-tumor T cell immunity, suggesting that PRMT1 inhibition may be an effective strategy to improve the efficacy of immune checkpoint blockade (ICB). [1]

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Epigenetic alterations are common events in clear cell renal cell carcinoma (ccRCC), and protein arginine methyltransferase 1 (PRMT1) is an important epigenetic regulator in cancer. However, the role of PRMT1 in clear cell renal cell carcinoma (ccRCC) remains unclear. Methods: We investigated the expression level of PRMT1 in CCRCC patients and its correlation with clinicopathological factors and prognosis using ccRCC tissue microarray (TMA). We employed gene knockdown and pharmacological inhibition with the novel PRMT1 inhibitor DCPT1061 to investigate the functional role of PRMT1 in ccRCC proliferation. Furthermore, we validated the antitumor effect of the PRMT1 inhibitor DCPT1061 in ccRCC cell-derived tumor xenograft (CDX) and patient-derived tumor xenograft (PDX) models. Results: We found that PRMT1 was significantly upregulated in tumor tissues and associated with adverse pathological features and prognosis in ccRCC patients. In addition, gene knockdown and pharmacological inhibition with the novel, potent inhibitor DCPT1061 significantly induced G1 phase cell cycle arrest and inhibited the growth of ccRCC cells. Mechanistically, RNA sequencing and further validation revealed that lipocarrier protein 2 (LCN2) is a secreted glycoprotein associated with tumorigenesis, a key regulator of clear cell renal cell carcinoma (ccRCC) growth, and a functional downstream effector molecule of PRMT1. PRMT1 deficiency leads to epigenetic silencing of LCN2 autocrine secretion, which in turn reduces the level of downstream phosphorylated AKT (p-AKT), and ultimately reduces the level of phosphorylated RB (p-RB) through neutrophil gelatinase-associated lipocarrier receptor (NGALR) and leads to cell growth arrest. In addition, the inhibition of PRMT1 by DCPT1061 not only inhibited tumor growth, but also enhanced the sensitivity of ccRCC to sunitinib treatment by attenuating the upregulation of the sunitinib-induced LCN2-AKT-RB signaling pathway. Conclusion: In summary, our study revealed the role of PRMT1-dependent epigenetic mechanisms in the regulation of ccRCC tumor growth and drug resistance, suggesting that PRMT1 may be a promising target for the treatment of ccRCC patients. [2]

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DCPT1061 is a research-grade small-molecule PRMT inhibitor for epigenetics and oncology research. It is available as a liquid formulation (10 mM in DMSO) with pack sizes ranging from 1 mL to 100 mg or more, and is typically stored at -20°C for long-term stability. It is classified as a histone methyltransferase inhibitor and is used in studies investigating the role of arginine methylation in cancer pathogenesis, as well as in drug discovery programs targeting PRMTs for therapeutic intervention. The compound's selectivity for PRMT1, PRMT6, and PRMT8 makes it a valuable tool for dissecting the specific contributions of these isoforms to oncogenic signaling, distinguishing it from pan-PRMT inhibitors that target a broader range of PRMT family members. For research use only, not for human therapeutic use.

C17H20CL2N2O

339.259502410889

338.0952

2289726-31-2

169494473

Colorless to light yellow liquid

4.1

1

3

7

22

319

0

ClC1C(=CC=CC=1OC1C=CC=CC=1CN(C)CCNC)Cl

GCURTAFSSBAXNO-UHFFFAOYSA-N

InChI=1S/C17H20Cl2N2O/c1-20-10-11-21(2)12-13-6-3-4-8-15(13)22-16-9-5-7-14(18)17(16)19/h3-9,20H,10-12H2,1-2H3

N'-[[2-(2,3-dichlorophenoxy)phenyl]methyl]-N,N'-dimethylethane-1,2-diamine

DCPT1061; DCPT-1061

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 :≥ 250 mg/mL (~736.90 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.)