Emulsion composition for chemoembolization and method for producing same

Abstract

The present invention relates to an emulsion composition for chemoembolization comprising a nanoparticle comprising a drug and a biocompatible polymer, a water-soluble contrast agent and a water-insoluble contrast agent, and a water-insoluble drug as well as an aqueous drug can be administered in a form of stable emulsion, and drugs are slowly released, thereby enhancing the effect of chemoembolization.

Claims

1. An emulsion composition for chemoembolization, comprising a polymer nanoparticle comprising a water-soluble anticancer agent, a water-insoluble anticancer agent, or a combination thereof as active ingredients, and an amphiphilic block copolymer comprising a hydrophilic (A) block and a hydrophobic (B) block; a water-soluble contrast agent; and a water-insoluble contrast agent, wherein the hydrophilic (A) block is one or more selected from the group consisting of monomethoxypolyethylene glycol, monoacetoxypolyethylene glycol, polyethylene glycol, copolymer of polyethylene and propylene glycol, and polyvinyl pyrrolidone, and the hydrophobic (B) block is one or more selected from the group consisting of polylactide, polyglycolide, polycaprolactone, polydioxan-2-one, copolymer of polylactide and glycolide, copolymer of polylactide and polydioxan-2-one, copolymer of polylactide and polycaprolactone and copolymer of polyglycolide and polycaprolactone; and the water-soluble contrast agent is iopamidol, and the water-insoluble contrast agent is one or more selected from the group consisting of poppy fruit-derived iodized oil, soybean-derived iodized oil and ethiodol, wherein the diameter of the polymer nanoparticle is 10 nm to 60 nm.

2. The emulsion composition for chemoembolization according to claim 1, wherein the number average molecular weight of hydrophilic (A) block or hydrophobic (B) block is 500 to 50,000 Da.

3. The emulsion composition for chemoembolization according to claim 1, wherein the weight ratio of the hydrophilic (A) block and hydrophobic (B) block is 2:8 to 8:2.

4. The emulsion composition for chemoembolization according to claim 1, wherein the polymer nanoparticle further comprises a polylactic acid derivative containing a terminal group of a carboxylic acid.

5. The emulsion composition for chemoembolization according to claim 4, wherein the polylactic acid derivative containing a terminal group of carboxylic acid is one or more selected from the group consisting of the compounds of Formulas 1 to 6:
RO—CHZ-[A].sub.n-[B].sub.m—COOM  [Formula 1] in the Formula 1, A is —COO—CHZ—; and B is —COO—CHY—, —COO—CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2— or —COO—CH.sub.2CH.sub.2OCH.sub.2; and R is a hydrogen atom, acetyl, benzoyl, decanoyl, palmitoyl, methyl or ethyl group; and Z and Y are independently a hydrogen atom, methyl or phenyl group; and M is Na, K, or Li; and n is an integer from 1 to 30; and m is an integer from 0 to 20;
RO—CHZ—[COO—CHX].sub.p—[COO—CHY′].sub.q—COO—CHZ—COOM  [Formula 2] in the Formula 2, X is a methyl group; and Y′ is a hydrogen atom or phenyl group; and p is an integer from 0 to 25, and q is an integer from 0 to 25, on the proviso that sum of p and q is an integer from 5 to 25; and R is a hydrogen atom, acetyl, benzoyl, decanoyl, palmitoyl, methyl or ethyl group; and M is Na, K, or Li; and Z is a hydrogen atom, methyl or phenyl group;
RO-PAD-COO—W-M′  [Formula 3] in the Formula 3, W-M′ is ##STR00004## and PAD is selected from the group consisting of D,L-polylactic acid, D-polylactic acid, polymandelic acid, copolymer of D,L-lactic acid and glycolic acid, copolymer of D,L-lactic acid and mandelic acid, copolymer of D,L-lactic acid and caprolactone and copolymer of D,L-lactic acid and 1,4-dioxan-2-one; and R is a hydrogen atom, acetyl, benzoyl, decanoyl, palmitoyl, methyl or ethyl group; and M is independently Na, K, or Li;
S—O-PAD-COO-Q  [Formula 4] in the Formula 4, S is ##STR00005## and L is -NR1- or -0-, wherein R1 is a hydrogen atom or C.sub.1-10 alkyl; and Q is CH.sub.3, CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.3, CH.sub.2CH.sub.2CH.sub.2CH.sub.3, or CH.sub.2C.sub.6H.sub.5; and a is an integer from 0 to 4; and b is an integer from 1 to 10; and M is Na, K, or Li; and PAD is selected from the group consisting of D,L-polylactic acid, D-polylactic acid, polymandelic acid, copolymer of D,L-lactic acid and glycolic acid, copolymer of D,L-lactic acid and mandelic acid, copolymer of D,L-lactic acid and caprolactone and copolymer of D,L-lactic acid and 1,4-dioxan-2-one. ##STR00006## in the Formula 5, R′ is -PAD-O—C(O)—CH.sub.2CH.sub.2—C(O)—OM, wherein PAD is selected from the group consisting of D,L-polylactic acid, D-polylactic acid, polymandelic acid, copolymer of D,L-lactic acid and glycolic acid, copolymer of D,L-lactic acid and mandelic acid, copolymer of D,L-lactic acid and caprolactone and copolymer of D,L-lactic acid and 1,4-dioxan-2-one, and M is Na, K, or Li; and a is an integer from 1 to 4.
YO—[—C(O)—(CHX).sub.a—O—].sub.m—C(O)—R—C(O)—[—O—(CHX′).sub.b—C(O)—].sub.n—OZ  [Formula 6] in the Formula 6, X and X′ are independently a hydrogen, alkyl having 1 to 10 carbon atoms or aryl having 6 to 20 carbon atoms; and Y and Z are independently Na, K, or Li; and m and n are independently an integer from 0 to 95 to meet 5<m+n<100; and a and b are independently an integer from 1 to 6; and R is —(CH2)k—, divalent alkenyl having 2 to 10 carbon atoms, divalent aryl having 6 to 20 carbon atoms or combination thereof, wherein k is an integer from 0 to 10.

6. The emulsion composition for chemoembolization according to claim 4, wherein the terminal group of carboxylic acid in the polylactic acid derivative is fixed with a divalent or trivalent metal ion.

7. The emulsion composition for chemoembolization according to claim 6, wherein the divalent or trivalent metal ion is selected from the group consisting of calcium (Ca.sup.2+), magnesium (Mg.sup.2+), barium (Ba.sup.2+), chrome (Cr.sup.3+), iron (Fe.sup.3+), manganese (Mn.sup.2+), nickel (Ni.sup.2+), copper (Cu.sup.2+), zinc (Zn.sup.2+) and aluminum (Al.sup.3+).

8. The emulsion composition for chemoembolization according to claim 1, wherein the viscosity of the water-soluble contrast agent or water-insoluble contrast agent is 10 to 40 mPa at 37° C.

9. The emulsion composition for chemoembolization according to claim 1, wherein the polymer nanoparticle is dissolved in the water-soluble contrast agent so that the concentration of anticancer agent is 1 to 40 mg/ml based on the total emulsion composition.

10. The emulsion composition for chemoembolization according to claim 9, wherein the volume ratio of the water-soluble contrast agent in which the polymer nanoparticle is dissolved and water-insoluble contrast agent is 1:2 to 1:10.

11. The emulsion composition for chemoembolization according to claim 1, wherein the viscosity of the emulsion composition is 20 to 50 mPa at 37° C.

Description

BRIEF DESCRIPTION OF DRAWINGS

(1) FIG. 1 shows the change in precipitation amount of a drug after storing a Paclitaxel polymer nanoparticle solution.

(2) FIG. 2 shows the drug release test result of the Paclitaxel polymer nanoparticle emulsion.

(3) FIG. 3 shows the change in size of cancer tissues after administering the Paclitaxel polymer nanoparticle emulsion and doxorubicin emulsion.

(4) FIG. 4 shows the viable tumor portion result after administering the Paclitaxel polymer nanoparticle emulsion, doxorubicin emulsion and Paclitaxel solution.

(5) FIG. 5 shows the AST change after TACE with the Paclitaxel polymer nanoparticle emulsion, doxorubicin emulsion and Paclitaxel solution.

(6) FIG. 6 shows the ALT change after TACE with the Paclitaxel polymer nanoparticle emulsion, doxorubicin emulsion and Paclitaxel solution.

MODE FOR INVENTION

(7) Hereinafter, the present invention will be described in detail by examples. However, the following examples are intended to illustrate the present invention only, and the present invention is not limited by the following examples.

<Example 1> Polymer Nanoparticle Preparation

(8) Paclitaxel 450 mg, mPEG-poly(D,L-lactide) 7600 mg and polylactic acid sodium salt 1642 mg were weighed and put into a round flask and completely dissolved in a small amount of dichloromethane, and then the solvent was completely removed at the temperature of 40° C. using a rotary evaporator. Purified water was added thereto and it was completely dissolved and sterile filtrated and the drug concentration was to be 20 mg/ml, and then it was lyophilized, thereby preparing a polymer nanoparticle.

<Example 2> Preparation of Solution of Nanoparticle and Water-Soluble Contrast Agent and Confirmation of Solubility

(9) To Pamiray 250™ (Dongkook pharmaceutical) as a water-soluble contrast agent, the Paclitaxel polymer nanoparticle prepared in Example 1 was dissolved to make the Paclitaxel concentration 20 mg/ml. The prepared solution was stirred at 200 RPM at the temperature of 37° C. and the precipitation amount of the drug over time was quantified, thereby confirming the change in solubility of the drug. The test solution was aliquoted 500μ each, and 0, 0.5, 1, 2, 3, 4 and 8 hours later, the test solution was filtrated (Millex HV, PVDF, 0.22 μm) and the drug concentration comprised in the nanoparticle in the filtrated solution was measured by HPLC. The HPLC measurement conditions were shown in the following Table 1.

(10) TABLE-US-00001 TABLE 1 Column poroshell 120 PFP, 4.6 mm × 150 mm, 2.7 μm Flow rate 0.6 ml/min Moving bed Acetonitrile/water (45/55, V/V) Detector and wavelength UV, 227 nm Column temperature 30° C.

(11) The change in the drug precipitation amount after storing the Paclitaxel nanoparticle solution at 37° C. was shown in FIG. 1, and it was confirmed that the solubility of the drug at the temperature of 37° C. by 2 hours after preparation was maintained to about 20 mg/ml.

<Example 3> Water-in-Oil (W/O) Emulsion Preparation and Stability Evaluation

(12) The solution of Example 2 and Lipiodol™ as the water-insoluble contrast agent were filled into a syringe respectively so that the volume ratio was 1:4, and then pumping was carried out using a 3-way stopcock 50 times or more, to prepare emulsion.

(13) In order to evaluate the stability of the prepared emulsion, the size change of droplets was observed by observing with an optical microscope at 25° C. over time. As a result, it could be confirmed that uniform droplets were formed in a size of 10 to 30 μm right after preparation.

(14) In addition, in order to confirm the amount of Paclitaxel which was moved to the water-insoluble contrast agent, Lipiodol layer, as the continuous phase in the prepared emulsion, by aliquoting 500 μ each and storing at the temperature of 37° C. for 1 hour while stirring, and then ultrafiltrating (molecular weight cut off: 100 kDa), the amount of the drug in the Lipiodol layer was HPLC analyzed under the analysis conditions of Table 1.

(15) The result was shown in FIG. 2, and it was confirmed that the Paclitaxel moved to the Lipiodol layer right after preparation and 1 hour later was less than 10% of the total.

<Example 4> Chemoembolization in Liver Cancer Animals

(16) 4.1 Animal Model Manufacturing

(17) After exposing left lobe of liver by performing laparotomy to Sprague Dawley rat, 1×10.sup.7 of N1-S1 rat hepatoma tumor cells (Seoul National University Hospital) were inoculated to manufacture a liver cancer animal model. 12 days later, MRI was carried out to confirm tumor formation and the size of tumor was measured.

(18) 4.2 Chemoembolization

(19) After the carotid artery of rat prepared in Example 4.1 was exfoliated and tied up with thread, thereby preventing hemostasis, the carotid artery was punctured using 24G medicut. Then, microguidewire and microcatheter were sequentially inserted. After implementing abdominal aorta angiography under induction of fluoroscopy, the emulsion of Example 3 was administered into liver artery, and doxorubicin as a control drug was dissolved in Pamiray 250™ as a water-soluble contrast agent at a concentration of 20 mg/ml, and the solution in which this and Lipiodol were mixed at a volume ratio of 1:4 (Pamiray:Lipiodol), and one in which Paclitaxel was dissolved in Lipiodol™ as a water-insoluble contrast agent were administered at the same dose. Then, in the computed tomography and magnetic resonance imaging, it was confirmed that Lipiodol was deposited inside the tumor, and after inserting a catheter through the carotid artery of the model mouse, chemoembolization was conducted.

(20) 4.3 Cancer Tissue Size Measurement

(21) one week, 2 weeks and 3 weeks later after conducting chemoembolization of Example 4.2, the size change of cancer tissue was measured using magnetic resonance, and the result was shown in FIG. 3. As shown in FIG. 3, it was confirmed that when administering the Paclitaxel polymer nanoparticle as emulsion using a water-soluble contrast agent and an oil contrast agent, the size of cancer tissue was not increased than doxorubicin emulsion.

(22) 4.4 Viable Tumor Portion Measurement

(23) On the 7th day after conducting TACE with the Paclitaxel polymer nanoparticle emulsion prepared in Example 2 and the doxorubicin emulsion and Paclitaxel solution as comparative examples, the normal liver tissue and cancer tissue were isolated by conducting biopsy. The isolated tissue was stained with hematoxylin and eosin and thereby the viable tumor portion was confirmed and the result was shown in FIG. 4.

(24) As shown in FIG. 4, it was confirmed that the viable tumor portion of the Paclitaxel polymer nanoparticle emulsion administration group was similar to the water-soluble anticancer agent, doxorubicin emulsion control group, despite of using a water-insoluble anticancer agent, and the tumor survival portion was low when comparing with the Paclitaxel solution-administered control group.

(25) 4.5 AST and ALT Measurement

(26) In order to confirm the hepatotoxicity, AST and ALT were measured by collecting blood before and 1, 3 and 7 days after TACE to the Paclitaxel polymer nanoparticle emulsion, doxorubicin emulsion and Paclitaxel solution.

(27) The measurement result was shown in FIG. 5 and FIG. 6, and it was confirmed that AST and ALT were significantly increased on the 1st day of TACE in all test groups, but they were almost completely recovered on the 3rd day. In particular, it was confirmed that the increase of AST and ALT on the 1st day of administration was relatively small, when administering the Paclitaxel nanoparticle emulsion formulation, compared to administering the Paclitaxel Lipiodol solution.