NOVEL LAPPACONITINE DERIVATIVE AND USE THEREOF
20240083850 ยท 2024-03-14
Assignee
Inventors
Cpc classification
A61K9/2018
HUMAN NECESSITIES
A61K9/2059
HUMAN NECESSITIES
C07D223/32
CHEMISTRY; METALLURGY
A61K47/26
HUMAN NECESSITIES
A61K9/2027
HUMAN NECESSITIES
International classification
C07D223/32
CHEMISTRY; METALLURGY
A61K9/48
HUMAN NECESSITIES
Abstract
The present invention pertains to a novel lappaconitine derivative, a method for producing same, and a pharmaceutical use thereof using osteogenesis-promoting activity. The lappaconitine derivative induces the differentiation of stem cells into osteoblasts, increases bone mineral density when administered to animal models of osteoporosis, and induces bone formation in animal models of bone fracture, and thus can be advantageously used for preventing, ameliorating, or treating bone-related diseases such as osteoporosis, as well as treating non-disease fractures caused by physical trauma.
Claims
1. A compound represented by the following Formula 1, and a stereoisomer, a hydrate, a solvate, or a pharmaceutically acceptable salt thereof: ##STR00006## wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, a C1-C6 alkyl, a thioalkyl, an arylalkyl, a hydroxyalkyl, a dihydroxyalkyl, a hydroxy alkylarylalkyl, a dihydroxyalkylarylalkyl, an alkoxyalkyl, an acyloxyalkyl, an aminoalkyl, an alkylaminoalkyl, an alkoxycarbonylaminoalkyl, an acylaminoalkyl, an arylsulfoneamidoalkyl, an allyl, a dihydroxyalkylallyl, a dioxolanylallyl, a caralkoxyalkyl, methyl, ethyl, isopropyl, t-butyl, and phenyl.
2. A pharmaceutical composition for preventing or treating a bone-related disease, comprising a compound represented by the following Formula 1, and a stereoisomer, a hydrate, a solvate, or a pharmaceutically acceptable salt thereof as an active ingredient: ##STR00007## wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, a C1-C6 alkyl, a thioalkyl, an arylalkyl, a hydroxyalkyl, a dihydroxyalkyl, a hydroxy alkylarylalkyl, a dihydroxyalkylarylalkyl, an alkoxyalkyl, an acyloxyalkyl, an aminoalkyl, an alkylaminoalkyl, an alkoxycarbonylaminoalkyl, an acylaminoalkyl, an arylsulfoneamidoalkyl, an allyl, a dihydroxyalkylallyl, a dioxolanylallyl, a caralkoxyalkyl, methyl, ethyl, isopropyl, t-butyl, and phenyl.
3. The pharmaceutical composition of claim 2, wherein the bone-related disease is selected from the group consisting of osteoporosis, bone fractures, rheumatoid arthritis, periodontitis, osteomalacia, osteopenia, bone atrophy, osteoarthritis, bone defects, osteolysis, and osteonecrosis.
4. The pharmaceutical composition of claim 2, wherein the compound represented by Formula 1 promotes the differentiation of stem cells into osteoblasts.
5. The pharmaceutical composition of claim 2, wherein the compound represented by Formula 1 increases the expression of an osteogenic marker selected from the group consisting of runt-related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), and osteocalcin.
6. A food composition for preventing or ameliorating a bone-related disease, comprising the compound of claim 1 as an active ingredient.
7. The food composition of claim 6, wherein the compound represented by Formula 1 promotes the differentiation of stem cells into osteoblasts.
8. A method of synthesizing the compound of claim 1, comprising: (a) allowing lappaconitine to react with an oxidizing agent; and (b) allowing the reaction product of (a) to react with an organic solvent in the presence of a base.
9. The method of claim 8, wherein the oxidizing agent of (a) is selected from the group consisting of phenyliodine diacetate (PhI(OAc).sub.2, lead (II) acetate (Pb(CH.sub.3CO.sub.2).sub.2), lead (II) acetate (Pb(CH.sub.3CO.sub.2).sub.4), ozone, and HIO.sub.4.
10. The method of claim 8, wherein the base of (b) is selected from the group consisting of sodium hydroxide, potassium carbonate, sodium carbonate, cesium carbonate, and potassium hydroxide.
11. The method of claim 8, wherein the organic solvent of (b) is an aliphatic alcohol or an alkoxy alcohol.
12. A method of treating a bone-related disease, comprising: administering the pharmaceutical composition of claim 2 to a subject in need of treatment.
Description
DESCRIPTION OF DRAWINGS
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BEST MODE
[0070] Hereinafter, one or more embodiments of the present invention will be described in detail with reference to examples thereof. However, it should be understood that the examples are for exemplary illustration and are not intended to limit the scope of the present invention.
Preparation Example: Preparation of QG3030
1-1. Preparation of Lappaconitine
[0071] Dichloromethane (500 mL) and a sodium hydroxide solution (10 g of NaOH, and 100 g of water) were added to lappaconitine.hydrogen bromide (10.08 g, 0.017 mol), and an organic layer and an aqueous layer were separated using a separatory funnel. The separated organic layer was washed several times with water, dried over anhydrous magnesium sulfate, and then distilled to obtain the target lappaconitine with a yield of 89% (8.90 g).
[0072] .sup.1H-NMR (CDCl.sub.3, 400 MHz)10.98 (s, 1H), 8.60 (d, J=8.4 Hz, 1H), 7.86-7.83 (m, 1H), 7.42 (t, 1H), 6.95 (t, 1H), 3.52-3.37 (m, 2H), 3.36-3.35 (m, 2H), 3.36 (s, J=7.1 Hz, 3H), 3.22 (s, 3H), 3.13 (s, 3H), 3.11-3.09 (m, 2H), 2.60-2.46 (m, 10H), 2.15 (s, 3H), 2.14-1.73 (m, 6H), 1.74-1.68 (m, 1H), 1.74-1.70 (m, 1H), 1.06 (t, J=6.8 Hz, 3H);
[0073] .sup.13C-NMR (CDCl.sub.3, 100 MHz) 169.02, 167.39, 141.6, 134.34, 131.07, 122.31, 120.19, 115.76, 90.10, 84.62, 84.13, 82.89, 78.57, 75.56, 61.48, 57.89, 56.52, 56.10, 55.50, 50.85, 49.86, 48.97, 48.51, 47.61, 44.76, 36.28, 31.83, 26.77, 26.20, 25.53, 24.12, 13.53;
[0074] HRMS (ES.sup.+): m/z calculated for C.sub.32H.sub.44N.sub.2O.sub.8: 585.3098 [M+H].sup.+.
[0075] Found 585.3176.
1-2. Preparation of Lappaconitine Derivative (LAD) from Lappaconitine
[0076] Lappaconitine (8.9 g, 0.015 mol) was slowly added to a solution in which phenyliodine diacetate (PhI(OAc).sub.2 (14.07 g, 0.044 mol) was dissolved in dimethylformamide (DMF, 150 mL), and stirred for 10 minutes. When the reaction was completed, the solution was diluted with ethyl acetate (EA), and extracted using an aqueous solution of saturated sodium bicarbonate (NaHCO.sub.3). The organic layer was washed several times with water to remove dimethylformamide, dried over anhydrous magnesium sulfate, and then distilled under reduced pressure. The resulting crude product was separated by column chromatography (diethyl ether:ethyl acetate:hexane=3:2:5) to obtain {(3S,6S,7S,9S,11S,16S)-1-ethyl-6,9,11-trimethoxy-8,13-dioxododecahydro-2H-3,6a,14-(epiethane[1,1,2]triyl)-7,10-methanocyclodeca[b]azocin-3(4H)-yl 2-acetaminobenzoate} as the target lappaconitine derivative (LAD) with a yield of 25.7% (2.3 g).
[0077] .sup.1H-NMR (CDCl.sub.3, 400 MHz)10.99 (s, 1H), 8.62 (d, J=8.4 Hz, 1H), 7.89 (d, J=9.6 Hz, 1H), 7.47-7.43 (m, 1H), 7.01-6.99 (m, 1H), 3.88-3.64 (m, 4H), 3.65 (s, 3H), 3.41 (s, 3H), 3.18 (s, 3H), 2.90-2.25 (5, 4H), 2.20 (s, 3H), 2.19-1.80 (m, 4H), 1.15 (t, J=7.0 Hz, 3H);
[0078] .sup.13C-NMR (CDCl.sub.3, 100 MHz) 211.85, 204.31, 169.10, 167.40, 162.18, 143.74, 141.74, 134.61, 131.07, 122.41, 120.30, 115.45, 86.92, 82.75, 81.45, 78.21, 76.28, 60.73, 58.17, 57.37, 54.35, 52.98, 50.19, 48.78, 46.06, 45.92, 38.41, 32.06, 25.56, 25.39, 25.36, 12.72;
[0079] HRMS (ES.sup.+): m/z calculated for C.sub.32H.sub.42N.sub.2O.sub.8: 583.3018 [M+H].sup.+.
[0080] Found 583.2941.
1-3. Preparation of QG3030 from LAD
[0081] LAD (2.3 g), a sodium hydroxide solution (0.7 g of NaOH and 5.1 g of water), and ethanol (51 mL) were put into a 100-mL 3-necked round bottom flask, and refluxed for an hour. When the reaction was completed, ethanol was distilled off, and the reaction mixture was subjected to column chromatography using ethyl acetate to obtain QG3030 {(2aR,2a1S,3S,4aS,8S,11S,11bS,12S)-6-ethyl-4a,8-dihydroxy-3,11-dimethoxy-tetradecahydro-8,11a,5-(epiethane[1,1,2]triyl)cyclopenta[7,1]indeno[5,4-b]azocin-1,2-dione} as the final compound with a yield of 62.5% (1.4375 g).
[0082] .sup.1H-NMR (DMSO-d.sub.6, 400 MHz)8.88 (s, 1H), 4.65 (s, 1H), 4.35 (s, 1H), 4.27-4.20 9m, 1H), 4.10-4.00 (m, 1H), 3.60 (s, 1H), 3.35 (s, 3H), 3.20 (s, 3H), 3.10-.2.99 (m, 1H), 2.85-2.19 (m, 6H), 1.99-1.50 (m, 7H), 1.05 (t, J=7.0 Hz, 3H);
[0083] .sup.13C-NMR (CDCl.sub.3, 100 MHz) 201.20, 149.88, 148.78, 83.25, 79.92, 79.50, 67.87, 62.74, 60.20, 58.10, 53.23, 49.55, 48.86, 47.88, 46.92, 42.39, 38.43, 13.56;
[0084] 108HRMS (ES.sup.+): m/z calculated for C.sub.22H.sub.31NO.sub.6: 406.2151 [M+H].sup.+.
[0085] Found 406.2242; X-ray structure.
1-4. Confirmation of Structure of QG3030
[0086] The QG3030 obtained in Preparation Example 1-3 was recrystallized with water, and appropriate crystals were obtained by X-ray diffraction. Thereafter, an X-ray structure of the crystals was determined (X-ray Diffractometer, R-AXIS RAPID). The results are shown in
TABLE-US-00001 TABLE 1 Molecular formula C.sub.22H.sub.31NO.sub.6 Molecular weight 405.49 Crystal Color, Habit Colorless, needle Crystal Dimensions 0.100 0.100 0.100 mm Crystal System Orthorhombic Lattice Type Primitive Lattice Parameters a = 8.864(3) .Math. b = 10.579(4) .Math. c = 20.680(7) .Math. V = 1939.1(12) .Math. .sup.3 Space Group P2.sub.12.sub.12.sub.1 (#19) Z value 4 D.sub.calc 1.389 g/cm.sup.3 F.sub.000 872.00 (MoKa) 1.003 cm.sup.1
Experimental Example 1: (In Vitro) Confirmation of Efficacy of QG3030
1-1. Confirmation of Osteocytic Differentiation Potential of QG3030 in MSCs
[0087] Osteoblasts involved in the osteogenesis are formed by differentiating mesenchymal stem cells (MSCs) using external stimuli and transcription factors involved in various signaling pathways (Garg et al., Orthop Surg. 2017). Accordingly, an experiment was performed to check whether QG3030 was able to induce the differentiation of human MSCs into osteoblasts.
[0088] MSCs from normal humans were purchased from the American Type Culture Collection (ATCC, U.S.A), and cultured in an MSC-specific culture medium (Gibco). The cultured cells were digested with trypsin, centrifuged, and then seeded in a 24-well plate at a concentration of 310.sup.4 cells/well. The next day, the MSC culture medium was exchanged with a DMEM (DMEM/10% FBS/penicillin/streptomycin) as an experimental medium, and the cells were treated with QG3030 alone (0.1 M) or OIM (osteogenesis-inducing medium; StemPro Osteogenesis Differentiation Kit, ThermoFisher Scientific Inc.,) as the positive control. Thereafter, the cell culture medium was exchanged with a fresh DMEM every day, treated with QG3030 or OIM, and then cultured for 3 weeks. After the culture was completed, the cells were stained with Alizarin Red for detection of calcium and Von Kossa for detection of mineral production.
[0089] Based on the staining results, it can be seen that QG3030 induced calcium and mineral production in human MSCs like the OIM as the positive control for osteoblast differentiation (
1-2. Confirmation of Change in Expression of RUNX2 and BMP2 in MSCs
[0090] Runt-related transcription factor 2 (RUNX2; core-binding factor alpha, Cbfa1) is the most important transcriptional regulatory factor for osteogenesis, and is known to be involved in osteoblast differentiation, matrix generation, and mineralization during osteogenesis (Bruderer M, et al., Eur Cell Mater. 2014). Also, RUNX2 itself may be regulated by a bone morphogenetic protein (BMP), which is a type of cytokine (Sun J, et al., Mol Med Rep. 2015). Accordingly, it was confirmed whether QG3030 promotes the expression of osteogenesis-related specific factors to induce osteogenesis.
[0091] MSCs were seeded in a 24-well plate at a concentration of 310.sup.4 cells/well, and cultured for 24 hours. From the next day, the MSCs were treated with different concentrations (0.001 M, 0.01 M, and 0.1 M) of QG3030 once a day for 7 days and 14 days. After the experiment was completed, the MSCs were stained with RUNX2 and BMP-2 antibodies, and observed using a fluorescence microscope.
[0092] As a result, it can be seen that QG3030 strongly induced the expression of RUNX2 and BMP-2 even at a low concentration (0.001 M=1 nM), and this tendency increased with an increasing concentration (
1-3. Confirmation of Change in Expression of Osteocalcin in MSCs
[0093] Osteocalcin is expressed only in osteoblasts (Lee et al., Cell 2007), and is used as a biomarker useful for osteogenesis (Bharadwaj et al., Osteoporosis International., 2009). Accordingly, it was confirmed that LAD had an effect on the expression of osteocalcin which is an osteoblast-specific marker.
[0094] An experiment was performed in the same manner as described in Experimental Example 1-2, and MSCs were then stained with an osteocalcin antibody, and observed using a fluorescence microscope. As a result, it was confirmed that QG3030 strongly induced the expression of osteocalcin within 7 days after the cells were treated with a low concentration (0.001 M) of QG3030, indicating that the expression level was similar to that of the OIM-treated group as the positive control (
1-4. Confirmation of Mechanism of Differentiation of MSCs into Osteoblasts
[0095] To study a mechanism by which QG3030 differentiates MSCs into osteoblasts, related signaling was investigated.
[0096] MSCs cultured in an experimental DMEM were treated with QG3030 (1 M) or the control compound (DMSO), and cultured for 24 hours. The cells were recovered, and an experiment was then performed according to a method specified in a phosphokinase antibody array kit (R&D Systems) containing antibodies that specifically recognize phosphorylated forms of 43 kinases. Then, samples were analyzed using an ImageJ (NIH, USA) program.
[0097] Based on the results of analysis, it can be seen that QG3030 strongly increased the phosphorylation of signaling molecules such as ERK, Akt, WNK1, and the like compared to the control DMSO (
1-5. Confirmation of Mechanism of Differentiation of MSCs into Osteoblasts
[0098] To further study the mechanism by which QG3030 differentiates MSCs into osteoblasts, an experiment was performed using a kinase inhibitor.
[0099] After MSCs were cultured, the MSCs were simultaneously treated with a kinase inhibitor (1 M) and QG3030 (1 M). The next day, the medium was exchanged, and the MSCs were simultaneously treated again with the kinase inhibitor and QG3030. This procedure was repeated for 3 weeks, and the MSCs were then stained with Alizarin Red.
[0100] As a result, it can be seen that the osteoblast differentiation ability of QG3030 was inhibited in the experimental group treated with a combination of QG3030 and the inhibitor, which inhibits ERK, P38, or Akt, compared to the QG3030-alone-treated group (
1-6. Confirmation of Effect on ERK Phosphorylation in MSCs
[0101] Previous research shows that an MAP-kinase signaling mechanism including ERK and p38 plays an important role in the osteoblast differentiation (Greenblatt et al., MB, Annu Rev Cell Dev Biol. 2013). Similar to the results of previous research, the results of Experimental Example 1-5 show that QG3030 shows the potential to differentiate MSCs into osteoblasts through ERK and p38 MAP-kinases (
[0102] As a result, it can be seen that the level of phosphorylated ERK (pERK) significantly increased within 30 minutes after QG3030 treatment, and specifically that the level of phosphorylated ERK (pERK) increased even after 24 hours after QG3030 treatment (FIG. 7A).
[0103] Also, to determine whether QG3030 induces ERK phosphorylation for a long period, MSCs were treated with various concentrations (0.001, 0.01, and 0.1 M) of QG3030, and the level of phosphorylated ERK (pERK) was checked after 7 days and 14 days. As a result, it was confirmed that the level of phosphorylated ERK (pERK) significantly increased in all the experimental groups treated with QG3030. In particular, it was confirmed that the level of phosphorylated ERK (pERK) increased to a level greater than that of the positive control (OIM) even at a low concentration. This tendency lasted until 14 days after QG3030 treatment, indicating that QG3030 increases ERK phosphorylation for a long period (
[0104] These results indicate that the long-term activation of an ERK signaling mechanism by QG3030 is likely to be a key mechanism of the osteoblast differentiation ability of QG3030.
1-7. Confirmation of Effect of ERK Inhibitor on Increase in RUNX2 Expression by QG3030
[0105] ERK may induce the phosphorylation of RUNX2 to increase transcriptional activity and may be involved in stability to increase an amount of RUNX2 protein (Greenblatt et al., MB, Annu Rev Cell Dev Biol. 2013). Therefore, an increase in ERK phosphorylation by QG3030 may induce RUNX2 expression so that QG3030 can be involved in the differentiation of MSCs into osteoblasts. To verify this, an expression level of RUNX2 was determined after MSCs were treated with a combination of QG3030 (0.1 M) and the ERK inhibitor PD98059 (50 M).
[0106] As a result of confirmation, it can be seen that the expression of RUNX2 significantly increased within 12 hours after QG3030 treatment, but the expression of RUNX2 significantly decreased when MSCs were treated with a combination of QG3030 and the ERK inhibitor PD98059. The tendency was the same even 24 hours after QG3030 treatment (
[0107] As expected, QG3030 treatment also increased a phosphorylation level of ERK in MSCs, and an increase in the phosphorylation level of ERK was inhibited by PD98059 treatment (
[0108] The results show that the induction of ERK activation by QG3030 is a key mechanism of osteogenic activity.
Experimental Example 2: (In Vivo) Confirmation of Efficacy of QG3030
2-1. Confirmation of Therapeutic Ability of QG3030 Using Animal Model of Osteoporosis
[0109] An ability of QG3030 to treat osteoporosis was confirmed in an animal model of ovariectomized (OVX) mice commonly used as an animal model of osteoporosis.
[0110] As the positive controls, parathyroid hormone (PTH)-based Forteo (Eli Lilly) used as an osteoporosis therapeutic agent through an osteogenesis-promoting mechanism, and alendronate-based Fosamax (Merck) serving as a bone resorption inhibitor were subcutaneously administered. As the negative control, water (H.sub.20) was orally administered, and QG3030 was orally administered at a concentration of 5 or 30 mg/kg (
[0111] As a result of measurement, it was confirmed that the bone mineral density significantly increased in the QG3030-administered groups compared to the negative control, indicating that QG3030 had a therapeutic effect on osteoporosis. In this case, the effect of QG3030 was similar to that of Fosamax, and significantly superior to that of Forteo (
[0112] Based on the tissue staining results, it can also be seen that QG3030 induced osteogenesis in the femurs of ovariectomized mice (
[0113] The experimental results are summarized in Table 2 below. In Table 2, BMD represents bone mineral density, TV represents total volume, BV represents bone volume, Tb.Th represents trabecular thickness, and Tb.N represents trabecular number.
TABLE-US-00002 TABLE 2 BMD TV BV BV/TV Tb. Th Tb. N Group (g/cm{circumflex over ()}3) (mm{circumflex over ()}3) (mm{circumflex over ()}3) (%) (1/mm) (mm) Sham 0.2303433 15.062852 1.225416 8.173108 0.078936 1.034056 OVX 0.1927733 14.942314 0.734676 4.959768 0.073672 0.670696 Fosamax 0.2135133 15.509586 1.017400 6.521050 0.069936 0.928632 Forteo 0.1997300 15.431426 0.879062 5.734686 0.071124 0.801822 QG3030-5 mg 0.2136333 16.571958 1.106886 6.712756 0.070814 0.947182 QG3030-30 mg 0.2300667 14.920258 1.155692 7.731468 0.074920 1.043926
2-2. Confirmation of Therapeutic Ability of QG3030 Using Animal Model of Bone Fracture
[0114] The ilium of a Sprague Dawley (SD) rat was subjected to drilling with a diameter of 2 mm to induce a bone defect, and QG3030 (6, 30, and 150 g/10 L) mixed with an excipient (collagen sponge, CollaCote) was implanted into a bone defect site to evaluate an osteoanagenic effect (
TABLE-US-00003 TABLE 3 Animal group Concentration Animal number Sham 5 Fracture + DMSO Collagen sponge + DMSO (10 uL) 5 Fracture + BMP2 Collagen sponge + BMP2 (1 ug/10 uL) 5 Fracture + QG3030 (low) Collagen sponge + QG3030 (6 ug/10 ul) 5 Fracture + QG3030 (middle) Collagen sponge + QG3030 (30 ug/10 ul) 5 Fracture + QG3030 (high) Collagen sponge + QG3030 (150 ug/10 ul) 5
[0115] Based on the Micro-CT image reading results 4 weeks after implantation, it can be seen that QG3030 shows a level of osteoanagenic capacity similar to BMP2 at all concentrations (
[0116] To statistically quantify the osteoanagenic capacity of QG3030, a degree of osteogenesis was realized with a 3-D image, and a region of interest (ROI) was set to analyze parameters such as bone volume/total volume (BV/TV; %), trabecular thickness (Tb.Th; mm), trabecular number (Tb.N; 1/mm), and trabecular separation (Tb.Sp; mm).
[0117] As a result, it can be seen that the bone volume/total volume (BV/TV) as a marker for osteogenesis increased to a similar level in the BMP2-treated group (positive control) and QG3030-treated groups (low, medium, and high concentrations) compared to the negative control treated with DMSO (
[0118] From the analysis of the trabecular thickness (Tb.Th), the trabecular number (Tb.N), and the trabecular separation (Tb.Sp) to evaluate the bone quality of the generated bone tissue, it was also confirmed through the Tb.N and Tb.Sp other than Tb.Th that the high-quality bone tissue was statistically significantly formed in the BMP2-treated group (positive control) and QG3030-treated groups (low, medium, and high concentrations) compared to the DMSO-treated group (
[0119] To verify the osteoanagenesis by QG3030 at the tissue level, tissue staining for cells (H&E, hematoxylin&eosin), collagen (Masson's trichrome), type I collagen, and osteocalcin was performed. As a result, it was confirmed that the markers were expressed at bone defect sites (indicated by solid lines) treated with QG3030 (low, medium, and high concentrations), and the effects was similar to that of BMP2 (
[0120] The results of osteoanagenesis in this experimental example verify that QG303O possesses the same excellent osteoanagenic capacity as BMP2.
[0121] Based on the results, it was confirmed that the QG3030 compound of the present invention induces calcium and mineral production in MSCs, increases the expression of RUNX2, BMP-2, and osteocalcin, and induces ERK activation to induce osteoblast differentiation. Also, because the QG3030 compound increases bone mineral density when administered to the animal model of osteoporosis and induces osteogenesis in the animal model of bone fracture, the QG3030 compound may be effectively used for treating non-disease bone fractures caused by physical trauma as well as the bone-related disease such as osteoporosis.
Formulation Examples
[0122] Meanwhile, the novel compound QG3030 according to the present invention may be formulated into various forms according to a purpose. Hereinafter, methods of formulating a composition including the novel compound QG3030 of the present invention as an active ingredient are described for illustrative purposes, but the present invention is not limited thereto.
1. Tablet (Direct Pressing)
[0123] 5.0 mg of the active ingredient was sieved, mixed with 14.1 mg of lactose, 0.8 mg of crospovidone USNF, and 0.1 mg of magnesium stearate, and then pressed to prepare tablets.
2. Tablet (Wet Assembly)
[0124] 5.0 mg of the active ingredient was sieved, and then mixed with 16.0 mg of lactose and 4.0 mg of starch. 0.3 mg of polysorbate 80 was dissolved in pure water, and a suitable amount of this solution was then added to the mixture for granulation. After drying, fine granules were sieved, and then mixed with 2.7 mg of colloidal silicon dioxide and 2.0 mg of magnesium stearate. The fine granules were pressed to prepare tablets.
3. Powder and Capsule
[0125] 5.0 mg of the active ingredient was sieved, and then mixed with 14.8 mg of lactose, 10.0 mg of polyvinyl pyrrolidone, and 0.2 mg of magnesium stearate. The mixture was filled into hard No. 5 gelatin capsules using a suitable device.
4. Injection
[0126] 100 mg of the active ingredient, 180 mg of mannitol, and 26 mg of Na.sub.2HPO.sub.4/H.sub.2O were dissolved in 2,974 mg of distilled water to prepare an injection.