KR20200038820A - Pharmaceutical composition for prevention and treatment of mixed lineage leukemia-rearranged leukemia comprising Massonianoside B(MA) as an active ingredient - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing and treating mixed lineage leukemia-rearranged leukemia (MLL-rearranged leukemia), comprising, as an effective component, an inhibitor of disruptor of telomeric silencing 1-like (DOT1L), massonianoside B (MA). The present inventors have conducted studies for treatment of leukemia, and have found that, when MLL-rearranged leukemia cells are treated with MA, which is a natural product inhibitor of DOT1L, MA inhibits the expression of genes involved in causing leukemia, via selective inhibition on H3K79 methylation, and selectively inhibits the proliferation of MLL-rearranged leukemia cells. Consequently, MA, which is discovered as a novel compound different from the molecular structures of existing DOT1L inhibitors, is expected to be utilized in the development of new DOT1L inhibitors and research on DOT1L and mechanism of pathogenesis of MLL-rearranged leukemia.

Description

Pharmaceutical composition for prevention and treatment of mixed lineage leukemia-rearranged leukemia comprising Massonianoside B (MA) as an active ingredient}

The present invention is a mixed lineage leukemia-rearranged leukemia (hereinafter referred to as 'mixed lineage leukemia-rearranged leukemia'), which comprises a disruptor of telomeric silencing 1-Like (DOT1L) inhibitor Massonianoside B (MA) as an active ingredient. MLL-leukemia) relates to a pharmaceutical composition for prevention and treatment.

Mixed lineage leukemia-rearranged leukemia (MLL-leukemia) differs in acute leukemia and gene expression patterns, more than 70% of acute lymphocytic leukemia in infants, and 35% of acute myeloid leukemia in infants Occupies the ideal. Due to the poor effectiveness of currently available treatments, the prognosis for MLL-leukemia patients is poor, and the development and application of specific treatments for MLL-leukemia patients is urgently required.

Meanwhile, methyltransferases (PMTs) are enzymes involved in several biological mechanisms and are attracting attention as a promising therapeutic target for tumors and other diseases. Among them, DOT1L (Disruptor of telomeric silencing 1-like) H3K79 (Histone H3 lysine 79) methyl group transferase has been shown to be involved in the development and progression of MLL-leukemia, and DOT1L selective inhibitors are receiving attention as a treatment for MLL-leukemia, DOT1L It is expected that the development of selective inhibitors will be the starting point for the study of pathological mechanisms of MLL and the development of therapeutic agents. However, the DOT1L inhibitors developed to date are S-adenosylmethionine (SAM) or S-adenosylhomocysteine (SAH) cofactor-like structures containing adenosine or diazaadenosine bases, and when the inhibitor enters the cell The enzymes related to the ribose structure are rapidly degraded to show adverse pharmacokinetic properties such as low oral bioavailability and inhibition of oral absorption. In addition, there are no related drugs that have succeeded in clinical trials and market entry.

Therefore, there is a need to study a new structure of the MLL-leukemia therapeutic agent by developing a new DOT1L selective inhibitor that deviates from the existing SAM / SAH-like DOT1L inhibitor and confirming cell-level activity.

Republic of Korea Patent Publication 10-2013-0125379

The present invention has been devised to solve the problems in the prior art as described above, and mixed linearity comprising Massonianoside B (MA), a DOT1L (Disruptor of telomeric silencing 1-Like) inhibitor, as an active ingredient. Provided is a pharmaceutical composition for the prevention and treatment of leukemia (MLL-leukemia) with leukemia gene rearrangement (mixed lineage leukemia-rearranged leukemia).

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned can be clearly understood by those skilled in the art from the following description. There will be.

In order to achieve the object of the present invention as described above, the present invention is a mixed lineage leukemia gene material containing a Masonianoside B (Massonianoside B; MA) which is a disruptor of telomeric silencing 1-Like (DOT1L) inhibitor. Provided is a pharmaceutical composition for preventing and treating mixed lineage leukemia-rearranged leukemia (MLL-leukemia).

In another embodiment of the present invention, the MA may inhibit the expression of H3K79 (Histone H3 lysine 79) methylation or leukemia-generating gene.

In another embodiment of the present invention, the H3K79 methylation may be methylation of the histone protein H3K79me1 or H3K79me2.

In another embodiment of the present invention, expression of the leukemia-generating gene HOXA9 (Homebox A9) or MEIS1 (Meis Hombox 1) can be suppressed.

In another embodiment of the present invention, the expression of the H3K79 methylation or leukemia-generating gene may be expressed by recruiting DOT1L by an MLL fusion partner.

In another embodiment of the present invention, the MLL fusion partner may include one or more partners selected from the group consisting of AF4, AF9, and AF10, and the carboxy terminal of the MLL fusion partner has an oncogenic fusion protein. prtein), and the carcinogenic fusion protein may include the amino terminal region of the MLL protein.

In another embodiment of the invention, the carcinogenic fusion protein can be induced by chromosomal translocation.

The present inventors conducted extensive studies for the treatment of leukemia, and through the selective inhibition of H3K79 methylation when treated with MLL-leukemia cells (MV4-11 cell line), the natural product-derived DOT1L inhibitor Masonianoside B (MA) was treated. It was confirmed that it inhibited the expression of leukemia-inducing gene and selectively inhibited the proliferation of MLL-leukemia cells. As a result of discovering a new class of compound MA different from the existing DOT1L inhibitor molecular structure, new DOT1L inhibitor development and DOT1L and MLL -It is expected to be used for research on mechanisms related to the development of leukemia.

Figure 1a is a diagram showing the molecular structure of the natural product-derived DOT1L inhibitor Masonicanoside B (Massonianoside B; MA).
1B is a diagram showing a graph of inhibitory activity of SAH (S-adenosylhomocysteine) and MA according to concentration.
1C is a diagram showing a study of the mechanism of action of MA through a graph between SAM and IC ₅₀ .
1d is a diagram showing a schematic diagram of inhibitory activity (IC ₅₀ ) of MA in various human methyl transferases.
Figure 2a is a diagram showing the results of immunoblotting according to Example 2-1 of H3K79me2 by concentration of MA and EPZ5676 compounds in MV4-11 cells.
Figure 2b is a diagram showing the results of analyzing the size of the H3K79me2 band (band) compared to Histone H3.
Figure 2c is a view showing the results of immunoblotting according to Example 2-1 using individual methylated lysine target antibodies after 10 μM MA treatment on MV4-11 cells.
Figure 2d is a diagram showing the results of HOXA9 and MEIS1 mRNA expression through qRT-PCR analysis according to Example 2-2 in MV4-11 cells treated with EPZ5676 (10 μM) and MA (15 μM) for 7 days.
Figure 3a is a diagram showing the change in the number of cells per day of MV4-11 cells according to the MA concentration.
Figure 3b is a diagram showing the change in cell number per day of MV4-11 cells and non-rearranged Jurkat cells according to the presence or absence of MA treatment.

The present inventors conducted extensive studies for the treatment of leukemia, and through the selective inhibition of H3K79 methylation when treated with MLL-leukemia cells (MV4-11 cell line), the natural product-derived DOT1L inhibitor Massonianoside B (MA) was treated. Inhibiting the expression of leukemia-inducing gene and selectively inhibiting the proliferation of MLL-leukemia cells, the present invention was completed by discovering a new class of compounds, MA, different from the existing DOT1L inhibitor molecular structure.

Hereinafter, the present invention will be described in detail.

In an embodiment of the present invention, in order to discover a DOT1L selective inhibitor of a new structure, as a result of constructing an in silico drug-specific molecular group according to Example 1-1 and screening a natural compound database in more than 7,000 laboratories, MA was discovered. , To confirm whether MA has selectivity with DOT1L, as a result of performing HotSpot methyl group transferase assay (PMT activity assay) according to Example 1-2, MA has very high selectivity compared to other methyl group transferases. It was confirmed that the bio-luminescent-based DOT1L inhibition assay according to Example 1-3 was performed to investigate the active mechanism of MA, and as a competitive inhibitor with MA, the DOT1L cofactor SAM It was confirmed that it works (see Example 1).

In another embodiment of the present invention, in order to confirm the methylation change of histone protein when treating MLL-leukemia cells MV4-11, MA, H3K79me1 in which DOT1L is involved as a result of immunoblotting according to Example 2-1 , It was confirmed that only H3K79me2 is selectively reduced, and qRT-PCR according to Example 2-2 was actively performed in MLL to confirm a change in gene expression related to MLL-leukemia when MA was treated with MV4-11. It was confirmed that expression of both HOXA9 and MEIS1 genes in which transcription occurs is reduced, and as a result of trypan blue staining analysis according to Example 2-3 to analyze cell proliferation and viability in a leukemia cell line, MA It was confirmed that the MLL-leukemia cell line has proliferation inhibitory activity only (see Example 2).

Accordingly, the present invention is a mixed lineage leukemia-rearranged leukemia with a mixed lineage leukemia gene rearrangement that includes a disruptor of telomeric silencing 1-Like (DOT1L) inhibitor Massonianoside B (MA) as an active ingredient. ; MLL-leukemia) provides a pharmaceutical composition for prevention and treatment.

The term "Disruptor of telomeric silencing 1-Like (DOT1L)" as used in the present invention is a histone H3K79 (S. cerevisiae) methyl group transfer enzyme and is a protein found in humans as well as other eukaryotic organisms. It is an epigenetic mark conserved in the eugenes of many eukaryotic organisms, and the methylation of H3K79 by DOT1L gradually increases along the aging process, and DOT1L acts as a biological clock to actively check the time. can do.

The term "mixed lineage leukemia gene rearrangement leukemia (MLL-leukemia)" as used in the present invention, accounts for about 75% of infant acute leukemia and appears in the form of acute lymphocytic and acute myeloid leukemia. MLL-leukemia has a poor prognosis among acute leukemias, and the 5-year disease-free survival rate of MLL acute lymphocytic leukemia is only 40%. In addition, MLL-leukemia is induced by chromosomal translocation and results in a carcinogenic fusion protein comprising the amino terminal region of MLL and the carboxy terminus of various fusion partners. Some of the MLL fusion partners, including AF4, AF9, and AF10, abnormally recruit DOT1L to specific genomic loci, leading to increased H3K79 methylation and transcriptional activation of leukemia-generating genes, including HOXA9 and MEIS1.

Thus, the MA can inhibit the expression of H3K79 (Histone H3 lysine 79) methylation and leukemia-generating genes, but is not limited thereto.

At this time, H3K79 methylation may be methylation for the histone proteins H3K79me1 or H3K79me2, and the leukemia-generating gene may be HOXA9 (Homebox A9) or MEIS1 (Meis Hombox 1), but is not limited thereto.

The term "H3K79 methylation" used in the present invention, in the case of lysine 79 of histone H3, is located inside the nucleosome rather than the amino terminus of histone unlike other histone methylation sites. In yeast, it is known that methylation mainly occurs in the euchromatin region, and methylation occurs in the coding region of an activated gene. The same pattern is observed in mammals, and an enzyme called DOT1L is involved in this process.

The term "HOXA9 (Homebox A9)" as used in the present invention, the homeobox gene encoding the transcription factor in vertebrate animals is usually found in clusters A, B, C, and D in four separate chromosomes, and the expression of these proteins Is spatially and temporally regulated during embryonic development, the HOXA9 gene is part of the A cluster of chromosome 7 and encodes a DNA-binding transcription factor that can regulate gene expression, morphogenesis and differentiation. This gene is very similar to the Drosophila abdominal B (Abd-B) gene, and certain translocation events that cause fusion between the HOXA9 and NUP98 genes are associated with the development of myeloid leukemia. Read-through transcription exists between the upstream genes HOXA9 and HOXA10 (homobox A10).

The term "MEIS1 (Meis Hombox 1)" used in the present invention is a gene required for hematopoiesis, megakaryotic cell lineage development, and vascular patterning. It is in the TALE ('3 amino acid loop extension') family of homeodomain-containing proteins. It encodes the belonging homeobox protein. In addition, when inducing myeloid leukemia, it may act as a cofactor of HOXA7 and HOXA9.

In addition, the expression of the H3K79 methylation or leukemia-generating gene may be expressed by recruiting and expressing DOT1L by an MLL fusion partner, but the MLL fusion partner is selected from the group consisting of AF4, AF9, and AF10 May include one or more, and the carboxy terminus of the MLL fusion partner may be included in an oncogenic fusion prtein, and the carcinogenic fusion protein may also include, but is not limited to, the amino terminal region of the MLL protein. Does not. In addition, the carcinogenic fusion protein may be induced by chromosomal translocation, but is not limited thereto.

The term "MLL fusion partner" as used in the present invention is a nuclear protein involved in the regulation of transcriptional elongation and the direct or indirect recruitment of the H3K79 histone methyl group transferase DOT1L. Many fusion partners, including cytoplasmic partners, have been found to form complexes with fusion proteins in the nucleus, and fusion partners also improve the stability of fusion proteins.

The term "oncogenic fusion prtein" used in the present invention is a protein produced by combining two or more genes that originally encode distinct proteins. This results in a single or multiple polypeptides with functional properties derived from each of the original proteins. Carcinogenic fusion proteins are usually naturally occurring fusion proteins, commonly found in cancer cells and can act as cancer proteins in cancer cells.

As used in the present invention, the term “chromosomal translocation” refers to a change in a chromosomal structure in which a chromosome fragment changes its position on another chromosome or on another site in the same chromosome. In addition to the chromosomal translocation, which is the cause of various genetic diseases and attached to other chromosomes of chromosomal fragments, where non-autosomal chromosomes exchange chromosomal fragments with each other, it may also involve deletion of the same chromosome and reverse translocation. Examples of actively using this broad translocation in gene expression in somatic cells include V (D) J recombination to form the variable region of the antigen receptor gene, and conversion recombination to replace the normal region of the immunoglobulin H chain gene. have. In addition, cancer genes such as c-myc and bcl-2 may translocate to the locus of the antigen receptor, causing lymphoma and lymphocytic leukemia.

As used in the present invention, the term “prevention” refers to all acts of inhibiting or delaying the onset of MLL-leukemia by preemptively administering the pharmaceutical composition according to the present invention before the onset of MLL-leukemia.

As used in the present invention, the term “treatment” refers to all actions in which symptoms of MLL-leukemia are improved or beneficially altered by administration of the pharmaceutical composition according to the present invention after the onset of mixed lineal MLL-leukemia.

Thus, suitable carriers, excipients, and diluents commonly used to prepare pharmaceutical compositions may be further included. In addition, it can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions according to a conventional method.

Carriers, excipients, and diluents that may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, and mineral oil. When formulating the composition, it is usually prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, surfactants.

The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "a pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type of patient's disease, severity, and activity of the drug , Sensitivity to drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs used, and other factors well known in the medical field.

The pharmaceutical composition according to the present invention, in order to enhance the therapeutic effect, preferably can be administered concurrently (simultaneous), separately (separate), or sequentially (sequential), and single or multiple administration of the combined drug . Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art. Specifically, the effective amount of the pharmaceutical composition according to the present invention may vary depending on the patient's age, sex, condition, weight, absorption of active ingredients in the body, inactivation rate and excretion rate, disease type, and drugs used in combination.

The pharmaceutical composition of the present invention can be administered to a subject by various routes. All modes of administration can be expected, for example, oral administration, intranasal administration, bronchoscopy, arterial injection, intravenous injection, subcutaneous injection, intramuscular injection, or intraperitoneal injection. The daily dosage is preferably divided into once to several times a day, but is not limited thereto.

The pharmaceutical composition of the present invention is determined according to the type of drug as the active ingredient, along with various related factors such as the disease to be treated, the route of administration, the patient's age, sex, weight, and severity of the disease.

Hereinafter, preferred embodiments are provided to help understanding of the present invention. However, the following examples are only provided to more easily understand the present invention, and the contents of the present invention are not limited by the following examples.

[Example]

Example 1. Excavation and identification of Massonianoside B (MA)

1-1. Pharmacophore-based screening

By constructing an in silico pharmacological group model with the structure and activity data of known DOT1L cofactor competitive inhibitors and screening the natural compound database in more than 7,000 laboratories, compounds containing the structural group at the adenosine base position were preferentially selected and finally Excavated as shown in Figure 1a.

1-2. DOT1L enzyme analysis and methyl transferase profiling (enzymatic assay and PMTs profiling)

As a result of comparing the activity between DOT1L and other representative methyl transferases (PMTs) by HotSpot methyl transferase activity assay (PMT activity assay), MA in DOT1L showed an IC ₅₀ of 383 ± 7.3nM level, and performed It showed a very high selectivity (more than 2000 times compared to PKMTs, more than 5000 times compared to PRMTs) compared to other methyl group transferases (FIGS. 1B and 1D, Table 1).

^a IC ₅₀ is ^a value obtained by performing a HotSpot PMT activity analysis by 'Reaction Biology Corporation' (Malvern, PA, USA).

^b fold selectivity is proportional to the DOT1L, it is calculated as the ratio of the IC ₅₀ of the IC ₅₀ for the enzyme dae DOT1L under study.

^c was measured in the PRC2 multiple protein complex.

^d was measured in PRMT5 / MEP50 complex.

1.3 Identification of the active mechanism of MA

Bioluminescent-based DOT1L inhibition assay was performed with Promega's Methyltransferase-Glo ^TM bioluminescent assay kit. Nucleosomes (Reaction biology HMT-35-130) purified from Hela cells were used as substrates, and all reactions were performed by diluting in methyltransferase assay buffer (total reaction volume: 20 μl). Active experiments were conducted with human recombinant DOT1L (residues 1-416, Reaction biology, HMT-11-101) 60 nM and Hela nuclear deficiency (oligonucleosome) at various concentrations of inhibitor candidates and SAM (125) ~ 10000nM). The reaction was divided in half into two wells in a white solid 96-well plate (Corning # 3992), reacted at 37 ° C. for 2 hours, and terminated with 0.1% TFA. Thereafter, MTase-Glo ^TM reagent and detection buffer were each treated in order, reacted for 1 hour, and then measured for luminescence. The inhibitory activity of the inhibitor candidates was measured by using wells not treated with the inhibitor candidate group as positive controls and wells not treated with inhibitor and DOT1L as negative controls.

As a result, as shown in Figure 1c, as the concentration of SAM (S-adenosylmethionine) increases, it was confirmed that the IC _{50 of} MA also increases, and it can be confirmed that MA acts as a competitive inhibitor with DOT1L cofactor.

Example 2. Identification of cellular activity of Masonianoside B (MA)

2-1. Confirmation of changes in histone protein methylation during MA treatment on MLL-leukemia cells MV4-11 using immunoblotting analysis

For Western blotting (Western blot = immunoblotting), MV4-11 cells were dispensed in a 6-well plate with 2 × 10 ⁵ cells per well with a total medium volume of 2 ml. Subsequently, DMSO control and inhibitors for each concentration were treated for 4 days, and histone proteins were obtained with a Total histone extraction kit (Epigenetic # 705010). Proteins were quantified in 10 μg increments, electrophoresed on a 15% Tris-glycine gel, and then transferred to a PVDF membrane (membrane) for 300 mA for 90 minutes. PVDF membrane (membrane) is then 0.1% TBST with 5% non-fat milk (blocking) in the condition until the next day after blocking (0.1% TBST) as a dilution buffer (dilution buffer) primary antibody for each manufacturer description After diluting as described above, the mixture was reacted at room temperature for 1 hour. The secondary antibody labeled with pepper peroxidase (Horseradish peroxidase labeled) was reacted for 1 hour at room temperature with a dilution ratio of 1: 4000 and the band was observed with ECL. The primary antibodies used are: H3K79Me1 (Abcam ab2886), H3K79me2 (Abcam ab3594), H3K9Me2 (Abcam ab1220), H3K9Me3 (Abcam ab6001), H3K27Me2 (Abcam ab110472), H3K27Me3 (Abcam ab108185) (Abcam ab108185) .

As a result, as shown in FIG. 2A, 1 μM EPZ5676 (positive control material that was administered in clinical phase 1 in MLL-leukemia patients) and 2 μM MA treatment showed significant H3K79me2 reduction in MV4-11 cells, as shown in FIGS. 2A and 2B. As shown in the figure, it was confirmed that H3K79me2 was reduced in a concentration-dependent manner with respect to MA, and it showed about 94% inhibition ability compared to the control at a concentration of 10 μM at H3K79me2 compared to the control histone H3. In addition, as shown in Figure 2c, it was confirmed that H3K79me1 is also reduced by MA treatment.

Furthermore, in order to confirm the selectivity of the methyl group transferase of MA, as a result of processing various methylated lysine target antibodies to MV4-11 cell extract protein treated with MA, MA did not affect lysine residues that DOT1L is not involved in ( H3K9, H3K27), and it was confirmed that only H3K79me1 and H3K79me2, which are involved in DOT1L, were selectively reduced. It was confirmed that MA was selective for DOT1L.

2-2. Confirmation of MLL-related gene expression change during MA treatment on MV4-11 cells using qRT-PCR method

The actively proliferating MV4-11 cells were dispensed in 2 ml of 2X10 ⁵ cells per well in a 6-well plate with a total medium volume of 2 ml. Subsequently, the RNA of the cells was purified with a Promega total isolation kit (Z3100) while replacing the medium every 3 to 4 days with 10 μM EPZ5676 and 15 μM RA conditions. Reverse transcription was performed using 1 μg of purified RNA using a high capacity cDNA reverse transcription kit (Applied biosystems 4368813), predesigned primer / probe set (HOXA9 (Hs00365956), MEIS1 (Hs00180020), TBP (4333769F), Applied biosystems), Taqman universal QPCR experiments were performed with PCR master mix (Applied biosystems 4304437).

Accordingly, as a result of performing qRT-PCR analysis on HOXA9 and MEIS1 genes where transcription is actively performed in MLL, as shown in FIG. 2D, expression of both HOXA9 and MEIS1 genes was reduced from 7 days after MA treatment on MV4-11 cells. Confirmed.

2-3. Cell proliferation and viability analysis

For the analysis of cell proliferation and viability in the human leukemia cell line, actively proliferating MV4-11 cells and control cells were dispensed in a 96-well plate with ³ × 10 ⁴ cells per well with a total medium volume of 150 μl. Subsequently, the inhibitor candidate groups were treated by concentration and the number of cells surviving for a total of 2 weeks at intervals of 3 to 4 days was measured according to a Trypan blue staining assay. The medium and the inhibitor were replaced every cell number measurement day, and the cell number was normalized to 5X10 ⁴ .

As a result, In the comparative experiments of cell proliferation and viability in the treatment of MA in MV4-11 cells and Jurkat control cells, MV4-11 cells showed a significant decrease in the number of cells in 12.5 μM MA conditions (FIG. 3A), but even higher concentrations in Jurkat cells No decrease in cell proliferation was observed (FIG. 3B). Through this, it was confirmed that MA has proliferation inhibitory activity only in the MLL-leukemia cell line.

The above description of the present invention is for illustration only, and those skilled in the art to which the present invention pertains can understand that it can be easily modified to other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and not restrictive.

Claims (9)

Mixed lineage leukemia-rearranged leukemia (MLL-rearranged leukemia) containing the disruptor of telomeric silencing 1-Like (DOT1L) inhibitor Massonianoside B (MA) as an active ingredient ) Preventive and therapeutic pharmaceutical composition.
According to claim 1,
The MA is H3K79 (Histone H3 lysine 79) characterized in that to inhibit the methylation or expression of leukemia-generating, pharmaceutical composition.
According to claim 2,
The H3K79 methylation is characterized in that the methylation of the histone protein H3K79me1 or H3K79me2, pharmaceutical composition.
According to claim 2,
The leukemia gene is characterized in that the HOXA9 (Homebox A9) or MEIS1 (Meis Hombox 1), pharmaceutical composition.
According to claim 2,
The expression of the H3K79 methylation or leukemia-generating gene is characterized in that the MLL fusion partner (MLL fusion partner) recruits and expresses DOT1L, a pharmaceutical composition.
The method of claim 5,
The MLL fusion partner, characterized in that it comprises at least one selected from the group consisting of AF4, AF9, and AF10, pharmaceutical composition.
The method of claim 5,
The MLL fusion partner is characterized in that its carboxy terminus is included in the oncogenic fusion protein (oncogenic fusion prtein), pharmaceutical composition.
The method of claim 7,
The carcinogenic fusion protein is characterized in that it comprises the amino terminal region of the MLL protein, pharmaceutical composition.
The method of claim 7,
The carcinogenic fusion protein is characterized in that induced by chromosomal translocation, pharmaceutical composition.

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