HYPERBRANCHED POLYESTER POLYOL DERIVATIVE AS DRUG SOLUBILIZER

Abstract

It is provided a composition comprising a hyperbranched polyester polyol derivative and a small molecule comprising at least one cyclic amine. The hyperbranched polyester polyol derivative is obtainable by a method comprising the following steps: a) reacting only glycidol and ε-caprolactone at a temperature lying in a range of between 40° C. and 140° C. to obtain a hyperbranched polyester polyol derivative in which caprolactone residues and glycerol residues are randomly arranged; b) reacting the hyperbranched polyester polyol derivative of step a) with a sulfation reagent to obtain a sulfated hyperbranched polyester polyol as hyperbranched polyester polyol derivative.

Claims

1. A composition comprising: i) a hyperbranched polyester polyol derivative obtainable by a method comprising the following steps: a) reacting only glycidol and c-caprolactone at a temperature lying in a range of between 40° C. and 140° C. to obtain a hyperbranched polyester polyol derivative in which caprolactone residues and glycerol residues are randomly arranged, b) reacting the hyperbranched polyester polyol derivative of step a) with a sulfation reagent to obtain a sulfated hyperbranched polyester polyol as hyperbranched polyester polyol derivative ii) and a small molecule comprising at least one amine group.

2. The composition according to claim 1, wherein the small molecule comprises a non-aromatic cyclic amine.

3. The composition according to claim 1, wherein the small molecule comprises an aliphatic amine.

4. The composition according to claim 1, wherein the small molecule has a pKa value of more than 5.

5. The composition according to claim 1, wherein the small molecule is a heterocyclic tumor signal transduction inhibitor comprising at least one cyclic amine.

6. The composition according to claim 1, wherein the small molecule is at least one chosen from the group consisting of M9831/VX-984, berzosertib, sunitinib, trametinib, dabrafenib, bortezomib, talazoparib, osimertinib, gefinitib, afatinib, erlotinib, lapatinib, neratinib, dacomitinib, bosutinib, dasatinib, imatinib, nilotinib, ponatinib, ibrutinib, cabozantinib, pazopanib, regorafenib, vemurafenib, rucaparib, olaparib, niraparib, selumetinib, entrectinib, idasanutlin, ipatasertib, lorlatinib, axitinib, glasdegib, gedatolisib, barasertib, encorafenib, binimetinib, cobimetinib, ruxolitinib, SAR405838, MI-773, APG-115, siremadlin, staurosporin, capivarsetib, uprosertib, GSK2110183, miransertib, BAY1125976, ravoxertinib, ulixertinib, fimepinostat, mocetinostat, belinostat, entinostat, alpelisib, GSK343, nedisertib, LY3023414, and S63845.

7. The composition according to claim 1, wherein the small molecule is at least one chosen from the group consisting of sunitinib, trametinib, dabrafenib, topotecan, idasanutlin, ruxolitinib, borussertib, and talazoparib.

8. The composition according to claim 1, wherein no covalent bond is formed between the hyperbranched polyester polyol derivative and the small molecule.

9. The composition according to claim 1, wherein the composition comprises an aqueous solvent.

10. A medical method for at least one of treatment and diagnosis of a person administering a composition of claim 1 to the person in need thereof.

11. The medical method of claim 10 for treating a tumor.

12. A method for manufacturing a composition according to claim 1, comprising the following steps: a) providing a small molecule in dry form or in form of an aqueous solution, wherein the small molecule comprises at least one amine group, b) adding an aqueous solution of the hyperbranched polyester polyol derivative to the small molecule, c) agitating the mixture obtained in the preceding step to obtain a composition of small molecule encapsulated by the hyperbranched polyester polyol derivative, and d) separating non-encapsulated small molecule from the composition of small molecule encapsulated by the hyperbranched polyester polyol derivative.

13. The method according to claim 12, wherein the separating step is performed by subjecting the mixture after step c) to at least one of a centrifugation and a chromatography.

14. The method according to claim 12, wherein the composition of small molecule encapsulated by the hyperbranched polyester polyol derivative is lyophilized after step d).

15. The method according to claim 12, wherein the method is carried out in no solutions other than aqueous solutions.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0086] Further details of aspects of the solution will be explained with respect to exemplary embodiments and accompanying Figures.

[0087] FIG. 1 shows the toxicity of sunitinib toward HeLa cells when comparing free sunitinib to sunitinib solubilized by the hyperbranched polyester polyol derivative as carrier.

[0088] FIG. 2 shows the toxicity of sunitinib toward KB-V1 cells when comparing free sunitinib to sunitinib solubilized by the hyperbranched polyester polyol derivative as carrier.

DETAILED DESCRIPTION

[0089] Standard Formulation Process

[0090] The encapsulation of the small molecule comprising at least amine group (drug) is conducted in aqueous medium, at room temperature. To the dry drug, in powder, an aqueous solution of a hyperbranched polyester polyol derivative (polymer) manufactured according to the procedure explained in WO 2019/096782 A1, freshly dissolved, is added. Typical setting: 10 mg drug, 100 mg polymer dissolved in 10 mL Milli-Q water. The solution is stirred at 1000 rpm, in dark, for 18 h. Afterwards the solution is centrifuged at 3000 rpm for 5 minutes. For colored drugs, the solution is additionally passed on a sephadex column. The solution is then lyophilized and stored at −20° C.

[0091] Sephadex Purification

[0092] For the separation of non-encapsulated drug from the solubilized (encapsulated) drug, the solution is passed on a sephadex column (size-exclusion chromatography), where the particles are separated on the base of the molecular weight. Sephadex: G25 (healthcare). Column length: ca. 20 cm. Depending on the volume and amount of formulation, the length can be adjusted (either reduced or expanded). Prior to utilization, the sephadex powder is swelled in water for at least 2 h. Generally, the sephadex is conserved in water to be ready to use.

[0093] Manufacturing of Waterless Formulation

[0094] The formulation is dissolved in a small volume of water, freeze-dried and hanged on the lyophilization instrument to remove all the water for at least 24 h.

Example #1: Sunitinib

[0095] The encapsulation is conducted according to the standard formulation process. After centrifuging the solution at 3000 rpm for 5 minutes, the supernatant is additionally passed on a sephadex column, G25. Column length: ca. 20 cm. The solution is then lyophilized and stored at −20° C.

Example #2: Dabrafenib

[0096] The encapsulation is conducted as in case of sunitinib.

Example #3: Trametinib

[0097] The encapsulation is conducted as in case of sunitinib.

[0098] pH Control Formulation Process

[0099] The encapsulation is conducted in aqueous medium, in acidic pH, at room temperature. HCl is added to the Milli-Q water to get a pH of 2. This solution is then used to dissolve the polymer and added to the dry drug. Typical setting: 10 mg drug, 100 mg polymer dissolved in 10 mL Milli-Q water. The solution is stirred at 1000 rpm, in dark, for 18 h. Afterwards the solution is centrifuged at 3000 rpm for 5 minutes. For colored drugs, the solution is additionally passed on a sephadex column. The solution is then lyophilized and stored at −20° C.

Example #4: Borussertib

[0100] The encapsulation is conducted according to the pH control formulation process. After centrifuging the solution at 3000 rpm for 5 minutes, the supernatant is recovered, lyophilized and stored dry at −20° C.

[0101] Quantification of the Drug Loading by Uv-Vis

[0102] For drugs presenting an absorption band at wavelength higher than 400 nm, UV-VIS analysis is chosen as quantification method. First, a calibration curve of the free drug in the suitable solvent mixture is recorded. Suitable combinations of solvent include mixtures of water and methanol, as well as of water and ethanol, in both cases in a ratio falling in a range of from 40 to 60% to 20 to 80%.

[0103] The calibration curve is prepared with at least 5 points, in a concentration range depending on the drug to be quantified. Linear regression is applied to generate the equation of the straight-line necessary to calculate the drug content.

[0104] The formulation is afterwards dissolved in the same medium and an UV-VIS spectrum is recorded. It is possible to use a straight-line equation to get the concentration of drug in solution. Knowing the total volume of the formulation, the content of drug, in mg, can be calculated.

[0105] The carrying potential (CP) of the polymer (solubilizing agent) is obtained as the ratio of the loaded and solubilized drug to the amount of solubilizing agent (Eq. 1).

[00001]$\mathrm{CP}=\frac{\mathrm{drug}\left(\mathrm{mg}\right)}{\mathrm{carrier}\mathrm{amount}\left(\mathrm{mg}\right)}*100$

[0106] Quantification of the Drug Loading by Elemental Analysis

[0107] For drugs presenting an absorption band at wavelength lower than 400 nm, the UV-VIS approach is not suitable, due to drug signals overlapping with signals of the solubilizing agent. Therefore, elemental analysis is chosen as alternative approach. The drugs chosen for the solubilization and entrapment contain nitrogen atoms, which are not present in the polymer structure. Via the quantification of the nitrogen content in the sample, it is possible to determine the amount of drug in it. For the loading quantification, the same equation is used as in the previous case.

[0108] The following tables (Table 5, Table 6 and Table 7) show the results of the performed analyses. In this context, h(PG-co-PCL).sub.60S.sub.0.95 was used as hyperbranched polyester polyol derivative. Its non-sulfated backbone has a molecular weight of 60 kDa. The polyester polyol derivative has a sulfation degree of 95%.

TABLE-US-00005 TABLE 5 Carrying potential of h(PG-co-PCL).sub.60S.sub.0.95 calculated by elemental analysis Polymer Drug Carrying amount amount potential Drug (mg) (mg) (wt %) Sunitinib 1.14 0.099 8.7%.sup.  Dabrafenib 508.71 8.0 1% Trametinib 58.8 1.2 2% Borussertib 1.62 0.197 12% 

TABLE-US-00006 TABLE 6 Carrying potential of h(PG-co-PCL).sub.60S.sub.0.95 calculated by UV Polymer Drug Carrying amount amount potential Drug (mg) (mg) (wt %) Sunitinib 236 25.54 11% M9831/VX-984 45 5 10% Berzosertib 45 5 10%

TABLE-US-00007 TABLE 7 Size and Z-potential investigation of h(PG-co-PCL).sub.60S.sub.0.95 - DLS in PB Solubilized Size Drug (volume) Z-potential Nude Polymer 12 ± 4 −48.9 Sunitinib 14 ± 6 −33.5 Dabrafenib 12 ± 1 −29.6 Trametinib 14 ± 2 −18.9

[0109] FIG. 1 shows the results of a first cell viability experiments in which the toxicity of free sunitinib was compared to that of sunitinib (small molecule comprising at least one amine group, namely a cyclic amine) encapsulated by h(PG-co-PCL).sub.60S.sub.0.95 (hyperbranched polyester polyol derivative). From these experiments, the IC.sub.50 values of free and encapsulated sunitinib for HeLa cells can be derived.

[0110] FIG. 2 shows the results of a second cell viability experiments in which the toxicity of free sunitinib was compared to that of sunitinib encapsulated by h(PG-co-PCL).sub.60S.sub.0.95. From these experiments, the IC.sub.50 values of free and encapsulated sunitinib for KB-V1 cells can be derived.

TABLE-US-00008 TABLE 8 IC.sub.50 values comparison of the drugs solubilized in DMSO and drugs solubilized in sulfated polyester polyols Compound Cell line IC.sub.50 (μg/mL) Sunitinib free drug Hela 3.523 h(PG-co-PCL).sub.60S.sub.0.95 @ sun Hela 5.685 Sunitinib free drug KB-V1 1.954 h(PG-co-PCL).sub.60S.sub.0.95 @ sun KB-V1 2.951