Method of killing osteosarcoma cells with bone cement formulation comprising calcium and lithium

Abstract

Use of a bone cement formulation in the manufacture of an anti-osteosarcoma medicament for killing osteosarcoma cells is provided, wherein the cement paste formulation contains a solid component having a lithium compound and a calcium compound.

Description

Technical Field

[0001] The present invention relates to the treatment of osteosarcoma with bone cement preparations comprising calcium and lithium, and in particular with bone cement pastes comprising calcium phosphate, calcium sulfate and a lithium compound. Background Art

[0002] Although adjuvant chemotherapy with extensive surgical treatment is clinically standardized for osteosarcoma treatment, however, the prognosis is poor with frequent lung metastases. Osteosarcoma is radioresistant to standard doses of radiation therapy, so radiation therapy is not used to treat this tumor type. However, although high-dose irradiation can be administered after extensive surgical intervention, postoperative radiation therapy harms normal tissue.

[0003] It is desirable to kill any possible residual osteosarcoma cells or inhibit their growth without adversely affecting normal tissues. Summary of the invention

[0004] An object of the present invention is to provide a method for killing osteosarcoma cells with a bone cement preparation after surgical treatment to remove osteosarcoma from a patient, which comprises implanting the bone cement preparation into a bone cavity formed by surgical treatment in the patient, wherein the cement paste preparation comprises a solid component, and the solid component comprises a lithium compound and a calcium compound.

[0005] Another object of the present invention is to provide a new use of a bone cement preparation in the manufacture of an anti-osteosarcoma drug for killing osteosarcoma cells.

[0006] Preferably, the cement paste formulation is an antimicrobial formulation disclosed in US Pat. No. 10,207,023 B2.

[0007] Preferably, the lithium compound comprises lithium carbonate (Li2CO3). BRIEF DESCRIPTION OF THE DRAWINGS

[0008] Figure 1 Results of viability testing of 143B osteosarcoma cells for CBCA-400 samples extracted for 24 hours, 48 ​​hours, and 72 hours (CBCA-24 hours, CBCA-48 hours, and CBCA-72 hours, respectively) are shown, with results of blank tests without CBCA-400 sample extraction also shown.

[0009] Figure 2 Results of viability testing of HOS osteosarcoma cells for CBCA-400 samples extracted for 24 hours, 48 ​​hours, and 72 hours (CBCA-24 hours, CBCA-48 hours, and CBCA-72 hours, respectively) are shown, with results of blank tests without CBCA-400 sample extraction also shown.

[0010] Figure 3 Results are shown for viability testing of MG 63 osteosarcoma cells for CBCA-400 samples extracted for 24 hours, 48 ​​hours, and 72 hours (CBCA-24 hours, CBCA-48 hours, and CBCA-72 hours, respectively), with results for blank tests without CBCA-400 sample extraction also shown. DETAILED DESCRIPTION

[0011] The bone cement preparation provided according to the present invention comprises a solid component, wherein the solid component comprises a lithium compound and a calcium compound.

[0012] Preferably, the bone cement formulation contains about 5-80%, more preferably about 10-70%, of the lithium compound based on the weight of the solid component.

[0013] Preferably, the lithium compound is a lithium salt, lithium oxide, lithium amide (LiNH2), lithium hydroxide or a lithium halide; and more preferably lithium carbonate, lithium sulfate, lithium phosphate, lithium oxide, lithium fluoride, lithium acetate, lithium bromide, lithium hydroxide, lithium nitrate, lithium nitrite, lithium iodide, lithium molybdate (Li2MoO4), lithium tetraborate (Li2B4O7), lithium citrate tetrahydrate (Li3C6H5O7·4H2O) or lithium stearate (LiC 18 H 35 O2); and most preferably lithium carbonate.

[0014] Preferably, the solid component is a powder component comprising a lithium compound and a calcium compound, wherein the calcium compound is selected from calcium phosphate, calcium sulfate, calcium oxide, calcium carbonate, calcium hydroxide, calcium magnesium phosphate, calcium nitrate, calcium citrate, calcium chloride and mixtures thereof; and more preferably the calcium compound is calcium sulfate, a calcium phosphate source or a mixture thereof. Preferably, the calcium phosphate is tetracalcium phosphate (TTCP), dicalcium phosphate, tricalcium phosphate, monocalcium phosphate or a mixture thereof. Preferably, the calcium sulfate is calcium sulfate hemihydrate (CSH), calcium sulfate dehydrate (CSD), anhydrous calcium sulfate or a mixture thereof.

[0015] When the solid component is a powder component, the bone cement formulation further comprises a setting liquid component in a ratio of liquid to powder of 0.20 ml / g to 0.80 ml / g.

[0016] The present invention also provides a method for treating a subject having osteosarcoma cells, comprising forming a bone cement paste by mixing a powder component and a solidifying liquid component of the bone cement preparation of the present invention; and injecting the bone cement paste into a hole or cavity in a bone of the subject, wherein the bone cement paste solidifies in the hole or cavity in need of the treatment.

[0017] Alternatively, the solid component is a block, or particles or fragments obtained by crushing the block. The block comprises a lithium compound and a calcium compound, wherein the calcium compound is selected from calcium phosphate, calcium sulfate, calcium oxide, calcium carbonate, calcium hydroxide, calcium magnesium phosphate, hydroxyapatite or a mixture thereof, and is preferably selected from tetracalcium phosphate, dicalcium phosphate, hydroxyapatite, calcium sulfate dihydrate, calcium sulfate hemihydrate or a mixture thereof.

[0018] Preferably, the block is formed by setting the bone cement paste in a mould.

[0019] The present invention also provides a method of treating a subject having osteosarcoma cells, comprising implanting the block, the particles or the fragments of the bone cement formulation of the present invention into a hole or cavity in a bone of the subject in need of the treatment.

[0020] experiment

[0021] Material:

[0022] CBCA-400 was prepared by mixing appropriate amounts of tetracalcium phosphate (TTCP), dicalcium phosphate (DCPA), calcium sulfate (CSH+CSD), and lithium carbonate (Li2CO3) powders in a weight ratio of 30.63:11.43:51.40:6.54.

[0023] The hardening (setting) solution used was 0.6 M diammonium hydrogen phosphate (NH4)2HPO4.

[0024] CBCA-400 and hardening solution are available from JOY MEDICAL DEVICES CORPORATION, 1F., No. 63, Luke 2nd Road, Luzhu District, Kaohsiung City, Taiwan, China.

[0025] CBCA-400 was mixed with the hardening solution at a ratio of 1 g:0.36 mL under agitation for one minute, and the resulting paste was formed into pellets by hand. The pellets were used for experiments immediately after preparation.

[0026] Osteosarcoma cell viability test

[0027] Cell viability testing was performed according to ISO 10993-5. An extraction method was used. The extracts were prepared by immersing the pellets in culture medium at a rate of 0.2 (g / ml) at 37°C for 24 hours, 48 ​​hours, and 72 hours, and then collecting the liquid by centrifugation. The cell concentration was 5×10 4Osteosarcoma cells in a medium with 5000 cells / ml were incubated in a 96-well microplate (100 μl / well, i.e., 5000 cells / well) at 37°C in a humidified atmosphere of 5% CO2. After 24 hours, the medium was aspirated, and the liquid (100 μl) collected by centrifugation as described above was added to the wells and incubated at 37°C for 48 hours. The liquid was aspirated, and a mixture of the medium (100 μl) and MTT (10 μl) was added to the wells and incubated at 37°C for 1 hour. Cell viability was measured by using an MTT assay. After incubation for 1 hour, the mixture of the medium and MTT was aspirated, and 100 μl DMSO was added to each cell of the 96-well microplate, and the absorbance (optical density, OD) at 590 nm was measured using a Multiskan FC Microplate Photometer.

[0028] Three different osteosarcoma cells were used in the tests;

[0029]

[0030] result

[0031] Figures 1 to 3 Results are shown for viability testing of three different osteosarcoma cells for CBCA-400 samples extracted for 24 hours, 48 ​​hours, and 72 hours (CBCA-24 hours, CBCA-48 hours, and CBCA-72 hours, respectively), with results for a blank test without CBCA-400 sample extraction also shown.

[0032] like Figures 1 to 3 As shown, the cell viability of three different osteosarcoma cells decreased significantly for 24-hour extraction, which was stable for 48-hour extraction and 72-hour extraction.

Claims

1. Use of a bone cement formulation in the manufacture of an anti-osteosarcoma implant for killing osteosarcoma cells in a patient, wherein the bone cement formulation is implanted into a bone cavity formed by surgical treatment for removing osteosarcoma in the patient, wherein the cement paste formulation comprises a solid component, the solid component comprising a lithium compound and a calcium compound.

2. The use according to claim 1, wherein the bone cement formulation contains 5-80% of the lithium compound based on the weight of the solid component.

3. The method according to claim 1, wherein the lithium compound is a lithium salt, lithium oxide, lithium amide (LiNH2), lithium hydroxide or lithium halide.

4. The use according to claim 1, wherein the lithium compound is lithium carbonate, lithium sulfate, lithium phosphate, lithium oxide, lithium fluoride, lithium acetate, lithium bromide, lithium hydroxide, lithium nitrate, lithium nitrite, lithium iodide, lithium molybdate (Li2MoO4), lithium tetraborate (Li2B4O7), lithium citrate tetrahydrate (Li3C6H5O7·4H2O) or lithium stearate (LiC 18 H 35 O2).

5. The use according to claim 1, wherein the lithium compound is lithium carbonate.

6. The use according to claim 1, wherein the solid component is a powder component comprising the lithium compound and the calcium compound, wherein the calcium compound is selected from calcium phosphate, calcium sulfate, calcium oxide, calcium carbonate, calcium hydroxide, calcium magnesium phosphate, calcium nitrate, calcium citrate, calcium chloride and mixtures thereof.

7. The use according to claim 6, wherein the calcium compound is calcium sulfate, calcium phosphate or a mixture thereof.

8. purposes according to claim 7, wherein the calcium phosphate is tetracalcium phosphate (TTCP), dicalcium phosphate, tricalcium phosphate, monocalcium phosphate or a mixture, and the calcium sulfate is calcium sulfate hemihydrate (CSH), calcium sulfate dehydrate (CSD), anhydrous calcium sulfate or a mixture thereof.

9. The use according to claim 6, wherein the bone cement preparation further comprises a solidifying liquid component having a liquid to powder ratio of 0.20 ml / g to 0.80 ml / g, and the powder component and the solidifying liquid component are mixed to form a paste, wherein the bone cement preparation implanted in the bone cavity comprises injecting the paste into the bone cavity, and the paste solidifies in the bone cavity.

10. The use according to claim 1, wherein the solid component is a mass, or particles or fragments obtained by breaking up the mass, wherein the mass is formed by solidifying the paste as claimed in claim 9.

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