Anti-multiple myeloma activity of rotigotine and uses

Abstract

The present application provides the anti-multiple myeloma activity and application of rotigotine. Experimental results show that rotigotine is relocated to the Tudor domain of the polycomb-like protein family PHF19 protein, and PHF19 is a significant marker of multiple myeloma. Rotigotine has good selectivity to non-polycomb-like proteins, effectively inhibits the viability of multiple myeloma cells MM1.S, and inhibits tumor growth in tumor-bearing mice, showing that rotigotine can become an anti-multiple myeloma drug.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and particularly relates to the anti-multiple myeloma activity and application of Rotigotine. BACKGROUND

[0002] Multiple myeloma is a hematopoietic system malignant tumor disease caused by abnormal proliferation of plasma cells in bone marrow, accounting for 10% of hematological malignancies. The median age of multiple myeloma is 65 years old, and its occurrence and development is a multi-step process during which a series of cytogenetic or genetic changes occur. High-risk myeloma patients account for about 25%, and are characterized by increased drug resistance, high mortality and early recurrence. At present, there is a lack of effective drug treatment. Therefore, discovering drugs for treating high-risk multiple myeloma patients has broad market value. SUMMARY

[0003] In recent years, Polycomb-like protein PHF19 has been identified as the most significant marker of high-risk multiple myeloma, and its Tudor domain is crucial to the occurrence and development of multiple myeloma. The present application screens and discovers that the old drug Rotigotine can bind to the Tudor domain of PHF19. Rotigotine is the English name of Rotigotine, which is a dopamine receptor agonist approved by FDA as a patch for the treatment of early Parkinson's disease. The present application finds that Rotigotine can be repositioned to PHF19, with an affinity of 3 μM, and the affinity of the homologous proteins PHF1 and MTF2 is 11 and 13 μM, respectively, and the selectivity to non-Polycomb protein family is more than 100 times, indicating that the possibility of off-target to non-Polycomb protein family and causing toxic side effects is low. Rotigotine effectively inhibits the viability of multiple myeloma cells MM1.S and inhibits the tumor growth of MM1.S tumor-bearing mice.

[0004] Specifically, the present application provides the following technical solutions:

[0005] In one aspect, the present application provides the use of Rotigotine in the preparation of a medicament for treating multiple myeloma.

[0006] In another aspect, the present application provides the use of a composition comprising Rotigotine in the preparation of a medicament for treating multiple myeloma.

[0007] In another aspect, the present application provides a method for treating multiple myeloma, which comprises administering an effective amount of Rotigotine or a composition comprising Rotigotine to a subject.

[0008] As used herein, the term "treatment" refers to any effect that beneficially improves or treats multiple myeloma by administering the composition of the present application.

[0009] The composition of the present application can contain or not contain a pharmaceutically acceptable carrier.

[0010] The composition of the present application can further comprise suitable excipients and diluents generally used for preparing compositions, such as pharmaceutical compositions. In addition, the composition of the present application can be prepared by general methods to be used as oral preparations (e.g., powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc.), external preparations, suppositories, or injections. As for suitable preparations known in the art, those listed in Remington’s Pharmaceutical Science (1985) can be used.

[0011] The composition of the present application can be provided in the form of an injection. For example, it can include subcutaneous injection, intramuscular injection, intravenous injection, intraperitoneal injection, epidural injection, or intrathecal injection, etc.

[0012] In some embodiments, the drug or pharmaceutical composition is administered to the subject by intraperitoneal injection.

[0013] In some embodiments, the pharmaceutical or pharmaceutical composition is in a package which further comprises a product insert which specifies that the amount of rotigotine is 5-60 mg / kg, for example 5-55 mg / kg, 5-50 mg / kg, 5-45 mg / kg, 5-40 mg / kg, 5-35 mg / kg, 5-30 mg / kg, 5-25 mg / kg, 5-20 mg / kg, 5-15 mg / kg, 5-14 mg / kg, 5-13 mg / kg, 5-12 mg / kg, 5-11 mg / kg, 5-10 mg / kg, 5-9 mg / kg, 5-8 mg / kg, preferably 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg, 23 mg / kg, 24 mg / kg, 25 mg / kg, 26 mg / kg, 27 mg / kg, 28 mg / kg, 29 mg / kg, 30 mg / kg, 31 mg / kg, 32 mg / kg, 33 mg / kg, 34 mg / kg, 35 mg / kg, 36 mg / kg, 37 mg / kg, 38 mg / kg, 39 mg / kg, 40 mg / kg, 41 mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg, 51 mg / kg, 52 mg / kg, 53 mg / kg, 54 mg / kg, 55 mg / kg, 56 mg / kg, 57 mg / kg, 58 mg / kg, 59 mg / kg, 60 mg / kg, more preferably 20 mg / kg or 40 mg / kg.

[0014] In some embodiments, the dosage of rotigotine is 5-60 mg / kg, for example 5-55 mg / kg, 5-50 mg / kg, 5-45 mg / kg, 5-40 mg / kg, 5-35 mg / kg, 5-30 mg / kg, 5-25 mg / kg, 5-20 mg / kg, 5-15 mg / kg, 5-14 mg / kg, 5-13 mg / kg, 5-12 mg / kg, 5-11 mg / kg, 5-10 mg / kg, 5-9 mg / kg, 5-8 mg / kg. Preferably, the dosage of rotigotine is 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, etc. 39 58 mg / kg, 59 mg / kg, 60 More preferably, the dosage of rotigotine is 20 mg / kg or 40 mg / kg.

[0015] In some embodiments, the subject is a mammal, including non-human primates, monkeys, cats, dogs, horses, rabbits, rodents (e.g., mice, rats, hamsters, and guinea pigs), cattle, sheep, goats, and humans. Preferably, the subject is a human.

[0016] In some implementations, the multiple myeloma is a high-risk multiple myeloma, i.e., the subject has an IgH translocation of chromosome 16q23 (c-MAF) or 20q11 (MAFB), t(14;16), t(14;20), or t(4;14).

[0017] definition

[0018] PHF19: PHD finger protein 19, also known as PCL3, is a member of the multicomb family of proteins. Its N-terminal Tudor domain specifically recognizes the trimethylation modification of histone H3K36.

[0019] Tudor domain: A conserved sequence of about 100 residues, mostly composed of 5 antiparallel β-sheets forming a barrel structure, and is one of the common domains mediating protein-protein interactions.

[0020] PHF1: PHD finger protein 1, also known as PCL1, is a member of the multicomb-like protein family. Its N-terminal Tudor domain specifically recognizes H3K36 trimethylation modification.

[0021] MTF2 (Metal-response element-binding transcription factor 2), also known as PCL2, is a member of the multicomb-like protein family. Its N-terminal Tudor domain specifically recognizes H3K36 trimethylation modification.

[0022] Polycomb group proteins (PcG proteins) are highly conserved epigenetic regulators named after a series of Drosophila mutant phenotypes. These proteins primarily function by forming two types of complexes: polycomb repressor complexes 1 and 2 (PRC1 / 2).

[0023] MM1.S: A high-risk multiple myeloma cell line with the genetic characteristic of IgH translocation on chromosome 16q23 (c-MAF) or 20q11 (MAFB).

[0024] RPMI 8226: a high-risk multiple myeloma cell line with genetic characteristics of t(14;16) or t(14;20).

[0025] U266: An intermediate-risk multiple myeloma cell line with a genetic characteristic of 11q13 chromosome (cyclin-D1) IgH translocation, accounting for approximately 15% of multiple myeloma cases.

[0026] NCL-H929: An intermediate to high-risk multiple myeloma cell line with the genetic characteristic t(4;14).

[0027] Binding affinity refers to the strength of the binding between a single biomolecule (e.g., protein or DNA) and its ligand / binding partner (e.g., drug or inhibitor). Binding affinity is generally determined by the equilibrium dissociation constant (K). dThis constant is used to measure and report the strength of bimolecular interactions and to rank such strengths. d The smaller the value, the greater the binding affinity of the ligand to its target. d The higher the value, the weaker the attraction and binding between the target molecule and the ligand.

[0028] Enthalpy: Represents the amount of heat released per mole of bound ligand, reflecting changes in hydrogen bonds and van der Waals bonds.

[0029] Equilibrium dissociation constant: i.e., K d , is the ratio of dissociation rate to binding rate, which reflects the affinity of a compound for a target.

[0030] Entropy: reflects changes in hydrophobic interactions and / or conformational changes.

[0031] Pan-inhibitor: An inhibitor that acts on two or more targets.

[0032] SMN: Survival motor neuron. The SMN complex catalyzes the assembly of small ribonucleoproteins (snRNPs), which are building blocks of the spliceosome and thus play an important role in the splicing of pre-mRNA in cells. The N-terminal Tudor domain of this protein recognizes symmetrical dimethylation of arginine.

[0033] TDRD3: Tudor domain-containing protein 3, which functions as a scaffold protein. Its Tudor domain specifically recognizes arginine dimethylation.

[0034] SND1: Staphylococcal nuclease domain-containing protein 1, which functions in the degradation of miRNAs, and its Tudor domain recognizes arginine dimethylation.

[0035] Chemical shift change: the change in chemical shift of each residue of a protein. 15 N and 1 The characteristic frequency of the H nucleus under a magnetic field is called chemical shift. Titration of small molecules can cause changes in the chemical shift of residues at the binding site.

[0036] IC 50 Half-inhibition concentration (WIC) refers to the concentration at which a substance achieves a 50% inhibitory effect on certain biological processes. In this invention, it refers to the inhibition of cell activity by a compound.

[0037] Non-comb-like protein family: The multicomb-like protein PHF19 has been identified as the most prominent biomarker for multiple myeloma. The non-comb-like protein family comprises numerous members with diverse functions. This invention discovered that rotigotine targets the Tudor domain of the multicomb-like protein family, but the drug needs to be selective to reduce off-target side effects. To conduct selectivity studies, this invention determined the affinity of rotigotine for multiple Tudor domains, including the affinity for the Tudor domain of the non-comb-like protein family, based on structural and sequence similarity. Attached Figure Description

[0038] Figure 1 The binding affinity of rotigotine to the Tudor domains of the multicomb-like family proteins PHF19, PHF1, and MTF2, as determined by isothermal titration calorimetry, is shown in Figure A. Figure A shows a binding affinity of rotigotine to the PHF19 Tudor domain of 3.0 μM, Figure B shows a binding affinity of rotigotine to the PHF1 Tudor domain of 11 μM, and Figure C shows a binding affinity of rotigotine to the MTF2 Tudor domain of 12.8 μM.

[0039] Figure 2 The binding affinity of rotigotine to the Tudor domains of non-comb-like family proteins SMN, TDRD3, and SND1, as determined by NMR chemical shift perturbation experiments, is shown. Figure A shows a binding affinity of rotigotine to the SMN Tudor domain of 0.4 mM, Figure B shows a binding affinity of rotigotine to the TDRD3 Tudor domain of greater than 0.3 mM, and Figure C shows a binding affinity of rotigotine to the SND1 Tudor domain of greater than 0.3 mM.

[0040] Figure 3 The effects of rotigotine on the activity of four different multiple myeloma cell lines, including RPMI1640, U266, NCL-H929, and MM1.S cells, were shown, with IC50 values. 50 The values ​​were 28, ND, 54, and 13 μM, respectively.

[0041] Figure 4 The effects of rotigotine on tumor weight and tumor volume in a mouse model of MM1.S cell tumors are shown. Figure A shows the change in mouse body weight over time at two different drug doses; Figure B shows the change in tumor weight over time at two different drug doses; Figure C shows the tumor size of five mice in each group after euthanasia at two different drug doses; Figure D is a statistical representation of Figure C. Data are presented as mean ± standard error, and p-values ​​were calculated using an unpaired Student's t-test (n=5). Detailed Implementation

[0042] To make the objectives, technical solutions, and advantages of the present invention clearer, the present invention will be further described in detail below with reference to specific embodiments and accompanying drawings.

[0043] Example 1: Determination of the binding affinity of rotigotine to human polycomb-like family proteins

[0044] First, gene fragments encoding human polycomb-like family proteins PHF19 Tudor (residues 38-95, SEQ ID NO:1), PHF1 Tudor (residues 28-87, SEQ ID NO:2), and MTF2 Tudor (residues 44-102, SEQ ID NO:3) were amplified and ligated into the pET28a vector (GE Heakthcare). The recombinant plasmids were transformed into *E. coli* BL21(DE3) competent cells. When the OD600 reached 0.8-1.0, 0.3 mM isopropyl thio-β-galactoside was added and the cells were cultured at 16°C to induce protein expression. After culturing for another 24 h, the cells were collected by centrifugation for protein purification. The binding affinity of rotigotine to PHF1, MTF2, and PHF19 Tudor proteins was determined using isothermal titration calometry (ITC) on a MicroCal PEAQ-ITC instrument at 25°C. Small molecule concentrations were quantified by acquiring one-dimensional spectra on a 500 MHz spectrometer. Protein samples were centrifuged to remove precipitates before the experiment and quantified using UV absorption (A280). Default experimental parameters were selected. The solution in the titration needle was added to the sample cell in 20 drops, with the first drop being 1 μL and the remaining 19 drops being 2 μL each. The titration needle stirring speed was 750 r / min; the reference energy was 5 μcal / s; the time interval between each two drops was 120 s; and the initial interval was 60 s. MicroCal PEAQ-ITC analysis software was used, and curve fitting was performed using a single-point binding mode to obtain the enthalpy and binding constant. The entropy change was calculated using the Gibbs free energy equation. Experimental results showed that rotigotine had a binding affinity of 3 μM for PHF19, and binding affinities of 12.8 and 11 μM for the MTF2 and PHF1 Tudor domains, respectively. Figure 1 Experimental results showed that rotigotine has no selectivity in binding affinity to multicomb-like family proteins. Therefore, rotigotine is a pan-inhibitor targeting multicomb-like family proteins.

[0045] Example 2: Determination of binding affinity of rotigotine to the Tudor domains of non-multicomb-like family proteins SMN, TDRD3, and SND1.

[0046] First, gene fragments encoding the Tudor domains of non-multicomb-like family proteins SMN (residues 91-151, SEQ ID NO:4), TDRD3 (residues 555-615, SEQ ID NO:5), and SND1 (residues 729-787, SEQ ID NO:6) were amplified and ligated into the pET28a vector (GE Heakthcare). The recombinant plasmids were then transformed into E. coli BL21(DE3) competent cells. 15 Nitrogen-labeled proteins are added during bacterial culture. 15 Using N-ammonium chloride as the nitrogen source, when the OD600 reached 0.8-1.0, 0.3 mM isopropyl thio-β-galactoside was added and the cells were cultured at 16°C to induce protein expression. After culturing the cells for another 24 h, the cells were collected by centrifugation for protein purification. The binding affinity of rotigotine to the SMN, TDRD3, and SND1 Tudor domains was determined using nuclear magnetic resonance chemical shift perturbation. 15 Nitrogen-labeled proteins SMN, TDRD3, and SND1Tudor were concentrated to 0.1 mM using a 5K ultrafiltration tube (Merck, UFC901096), stored in phosphate solution, and their two-dimensional spectra were acquired using a 500 MHz Varian spectrometer. The rotigotine molecular weight / protein molar ratio was set to 0:1–3:1, and the dissociation constant K0 was set to 0.1 mM. d Fit according to the following formula, P t and L t Δδ represents the total concentration of protein and ligand, respectively. obs Δδ represents the observed chemical shift perturbation relative to the free state of the protein. obs Defined as δ H and δ N The chemical shift changes in the direct and indirect dimensions are respectively represented by the maximum chemical shift perturbation Δδ. max and K d It was obtained by fitting the stoichiometric chemical shift change.

[0047]

[0048] Experimental results show that rotigotine has a binding affinity of 0.4 mM for the SMN Tudor domain, while its binding affinity for TDRD3 and SND is greater than 0.3 mM. Figure 2 Rotigotine, as a high-affinity inhibitor of the polycomb-like family protein PHF19, has more than 100 times the selectivity for non-polycomb-like family proteins, reducing the probability of off-target effects of the drug in cells and thus reducing toxic side effects.

[0049] Example 3: Effect of rotigotine on the activity of different multiple myeloma cell lines

[0050] Cell viability was measured using CellTiter-Glo luminescence assay. This invention tested the effect of rotigotine on the viability of four different multiple myeloma cell lines: RPMI 8226 (Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd., ZM0903), U266 (Shanghai Zhongqiao Xinzhou Biotechnology Co., Ltd., ZM0619), NCL-H929 (Nanjing Kebai Biotechnology Co., Ltd., CBP60243), and MM1.S (Shanghai Fuheng Biotechnology Co., Ltd., FH0714). The specific procedure was as follows: cell lines were cultured in RPMI 1640 + 10% fetal bovine serum + 1% streptomycin / penicillin at 37ºC with 5% CO2. 3 Cells were seeded into 96-well plates at a volume of 95 μl, with 5 μl of DMSO (as a control) or rotigotine molecule added simultaneously for treatment. Final concentrations were set at 0.125, 0.25, 0.5, 1, 2, 4, 8, 16, and 32 μM, with 6 replicates per group. After 72 h of incubation, an equal volume of CellTiter-Glo assay reagent was added, vortexed for 5 minutes, and incubated at room temperature for 30 minutes. The luminescence value was measured using a microplate reader. Cell viability was calculated using the following formula: Cell viability (%) = (RLU) / (RLU) 实验组 -R0LU 空白对照 ) / (RLU DMSO -RLU 空白对照 RLU stands for Relative Light Unit. Results showed that rotigotine could inhibit the activity of various multiple myeloma cell lines, exhibiting good inhibitory effects on high-risk MM cell lines MM1.S and RPMI-8226, with an IC50 concentration of [missing value]. 50 The inhibitory IC50 values ​​for intermediate-risk MM cell line NCL-H929 were 13 μM and 28 μM, respectively. 50 54 μM ( Figure 3 It had no significant inhibitory effect on the activity of low-risk MM cell line U266.

[0051] Example 4: In vivo efficacy evaluation of rotigotine in an MM1.S cell subcutaneous xenograft tumor model

[0052] To evaluate the in vivo efficacy of rotigotine in a subcutaneous xenograft tumor model of MM1.S cells, the specific procedure was as follows: MM1.S cells were cultured in 1640 + 10% FBS + 1% streptomycin / penicillin at 37℃ with 5% CO2. When the cell saturation reached 80-90%, the cells were harvested and counted. 0.2 ml (2×10⁻⁶ cells) was then transferred to the culture medium. 6MM1.S cells and matrix gel (manufacturer Corning, catalog number 354234) were mixed at a 1:1 (v:v) ratio and subcutaneously injected into the right posterior back of each CB17 SCID female mouse (these mice were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd.; SCID mice have a recessive gene mutation on chromosome 16, lacking functional T and B lymphocytes, 6-8 weeks old, weighing 18-20 g), with five mice per group. Subcutaneous tumor bearing status was observed after injection. Animals were weighed before grouping, and the tumor was considered complete when the average tumor volume reached approximately 80-100 mm. 3 Three groups were initially administered rotigotine via intraperitoneal injection once daily at doses of 20 mg / kg and 40 mg / kg (using saline as the solvent). Tumor volume and mouse weight were measured three times weekly using calipers. Mice were euthanized after 14 days of administration, and tumor volume and weight were then assessed. The results showed that at both 20 mg / kg and 40 mg / kg doses, the weight loss in the rotigotine-treated groups was less than 20% compared to the control group. Therefore, rotigotine was considered to have no significant effect on mouse weight, but it significantly inhibited tumor growth in MM1.S tumor-bearing mice, reducing tumor volume to 33% and 23% of the control group, and tumor weight to 36% and 25% of the control group, respectively. Figure 4 ).

[0053] sequence

[0054] SEQ ID NO:1 PHF19 Tudor domain amino acid sequence

[0055] SKLTEGQYVLCRWTDGLYYLGKIKRVSSSKQSCLVTFEDNSKYWVLWKDIQHAGVPGE

[0056] SEQ ID NO:2 PHF1 Tudor domain amino acid sequence

[0057] RPRLWEGQDVLARWTDGLLYLGTIKKVDSAREVCLVQFEDDSQFLVLWKDISPAALPGEE

[0058] SEQ ID NO:3 MTF2 Tudor domain amino acid sequence

[0059] CKFEEGQDVLARWSDGLFYLGTIKKINILKQSCFIIFEDSSKSWVLWKDIQTGATGSGE

[0060] SEQ ID NO:4 SMN Tudor domain amino acid sequence

[0061] QWKVGDKCSAIWSEDGCIYPATIASIDFKRETCVVVYTGYGNREEQNLSDLLSPICEVANN

[0062] SEQ ID NO:5 TDRD3 Tudor domain amino acid sequence

[0063] MWKPGDECFALYWEDNKFYRAEVEALHSSGMTAVVKFIDYGNYEEVLLSNIKPIQTEAWEE

[0064] SEQ ID NO:6 SND1 Tudor domain amino acid sequence

[0065] APRRGEFCIAKFVDGEWYRARVEKVESPAKIHVFYIDYGNREVLPSTRLGTLSPAFSTR

[0066] The specific embodiments described above further illustrate the purpose, technical solution, and beneficial effects of the present invention. It should be understood that the above descriptions are merely specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. Use of rotigotine in the preparation of a medicament or pharmaceutical composition for the treatment of multiple myeloma in a subject.

2. The use according to claim 1, characterized in that, The drug or drug composition is provided in the form of an injection.

3. The use according to claim 2, characterized in that, The injectable is intended for subcutaneous, intramuscular, intravenous, intraperitoneal, epidural, or intrathecal injection.

4. The use according to claim 3, characterized in that, The injectable is for intraperitoneal injection.

5. The use according to any one of claims 1-4, characterized in that, The dosage of rotigotine is 5-60 mg / kg.

6. The use according to any one of claims 1-4, characterized in that, The dosage of rotigotine is 5-55 mg / kg, 5-50 mg / kg, 5-45 mg / kg, 5-40 mg / kg, 5-35 mg / kg, 5-30 mg / kg, 5-25 mg / kg, 5-20 mg / kg, 5-15 mg / kg, 5-14 mg / kg, 5-13 mg / kg, 5-12 mg / kg, 5-11 mg / kg, 5-10 mg / kg, 5-9 mg / kg, or 5-8 mg / kg.

7. The use according to any one of claims 1-4, characterized in that, The dosage of rotigotine is 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 11 mg / kg, 12 mg / kg, 13 mg / kg, 14 mg / kg, 15 mg / kg, 16 mg / kg, 17 mg / kg, 18 mg / kg, 19 mg / kg, 20 mg / kg, 21 mg / kg, 22 mg / kg、23mg / kg、24mg / kg、25 mg / kg、26 mg / kg、27 mg / kg、28 mg / kg、29 mg / kg、30 mg / kg、31 mg / kg、32 mg / kg、33 mg / kg、34 mg / kg、35 mg / kg、36 mg / kg、37 mg / kg、38 mg / kg、39 mg / kg、40mg / kg mg / kg, 42 mg / kg, 43 mg / kg, 44 mg / kg, 45 mg / kg, 46 mg / kg, 47 mg / kg, 48 mg / kg, 49 mg / kg, 50 mg / kg, 51 mg / kg, 52 mg / kg, 53 mg / kg, 54 mg / kg, 55 mg / kg, 56 mg / kg, 57mg / kg, 58 mg / kg, 59 mg / kg or 60 mg / kg.

8. The use according to any one of claims 1-4, characterized in that, The dosage of rotigotine is 20 mg / kg or 40 mg / kg.

9. The use according to any one of claims 1-4, characterized in that, The subjects were mammals, including non-human primates, cats, dogs, horses, rabbits, rodents, cattle, sheep, goats, and humans.

10. The use according to any one of claims 1-4, characterized in that, The subjects were humans.

11. The use according to claim 9, characterized in that, The rodents are mice, rats, hamsters, or guinea pigs.

12. The use according to any one of claims 1-4, characterized in that, The multiple myeloma is a high-risk type of multiple myeloma, meaning that the subject has an IgH translocation on chromosome 16q23 (c-MAF) or 20q11 (MAFB), t(14;16), t(14;20), or t(4;14).

Images