Patent Application
20230248642
This invention discloses an injectable nanocomposite gel composition and the method of making the composition, which can be used as a vehicle to carry and deliver an active ingredient.
Injectable hydrogels have been considerably reported over decades in literature for a number of biomedical applications ranging from fillers, implantable vehicles, carrier for drugs, cells, and supplements, etc. Natural polysaccharides such as chitosan, alginates, hyaluronates, glycan, dextran, etc. have been received large attention in synthesis of specific hydrogel for medical application, due to their excellent biocompatibility, biodegradability, processability, and ease of chemical modification. Therefore, use of natural polysaccharides, either as a primitive form or as a modified form, such as hydrophobically-modified or amphiphilically-modified, had received enormous interests for medical uses. For drug delivery application, such modified version is able to form nano-size particles which can be used to entrap pharmaceutically active ingredients of different physicochemical properties (e.g., hydrophobic and hydrophilic properties) simultaneously, followed by controlled delivery, via vein administration, intramuscular injection, intraperitoneal injection or subcutaneous injection to the host for therapeutic purpose.
Injection of hydrogel will lead to the formation of a “depot” at the site of administration that slowly and continuously releases the drug to the tumor or diseased site and its surrounding tissue. This kind of injectable gel for physical targeting provides a number of advantages over passive or other actively targeted therapies in that it can deliver a drug throughout the tumor or diseased sites regardless of vascular status and/or biological environment surrounding the site of administration, thus providing accurate dosing without systemic toxicity or due to possible variants between genders, ages, and races. For instance, poloxamer gels have been widely applied in drug delivery since they are relatively easy to manufacture and already widely employed in the pharmaceutical industries as “generally regarded as safe” (GRAS) excipients. This type of hydrogels mainly focuses on poloxamer 407. For localized cancer therapy, intratumoral, peritumoreal, and intravesical injection of such type of hydrogel composed of Pluronic® F127 (F127) has been reported (Y. L. Lo, C. Y. Hsu, H. R. Lin, pH- and thereto-sensitive pluronic/poly(acrylic acid) in situ hydrogels for sustained release of an anticancer drug, J Drug Target, 21 (2013) 54-66). However, such poloxamer gels for drug delivery applications have substantial drawbacks including the gelation time being too long, poor stability, poor mechanical properties and short residence times due to rapid dissolution once placed in a biological environment.
US 20120100103 discloses an in situ-forming injectable hydrogel comprising two or more homogeneous or heterogeneous polymers, which are bonded to each other by a dehydrogenation reaction between phenol or aniline moieties on adjacent polymers. US 20140065226 provides compositions including an environmentally-responsive hydrogel and a biocompatible monomer or polymer including an amino acid side chain (i.e., having an amino acid linked to the remainder of the monomer or polymer through its side chain), which has environmentally-responsive behavior at physiological condition, such as temperature and is useful as injectable and topical formulations, particularly for biomedical applications such as so localized drug delivery.
US20150366975A1 discloses a thermosensitive injectable hydrogel based on hyaluronic acid and a copolymer of polyethylene oxide (PEO) and polypropylene oxide (PPO), which has a gel formation temperature from 30° C. to 37° C. The thermosensitive injectable hydrogel provides a potential drug delivery system that can increase therapeutic efficacy of the drug.
It is desirable to develop a new drug delivery system used for injection administration.
Accordingly, the present invention provides a new approach to deliver one or more active ingredients or drugs in humans by combining such amphiphilic nanoparticles with a self-sustained porous matrix phase to form a drug-carrying injectable nanocomposite hydrogel in either highly-viscous or solid form for a variety of medical uses, for example for anti-tumor treatment.
The present invention generally relates to an injectable nanocomposite gel composition and the method for preparing the same. In particular, the present invention relates to an injectable hydrogel.
In one aspect, the present invention provides a composition of injectable nanocomposite gel, which comprises an amphiphilic alginate nanoparticle, a hyaluronic salt or derivative, an alginate salt or derivative, and an ionic crosslinker.
In one embodiment of the invention, the composition further comprises an active ingredient. The active ingredient is selected from the group consisting of an antibody drug, a biosimilar drug, a protein-like drug, a chemo-drug, and the combination thereof.
In other embodiment of the invention, the active ingredient is selected from the group consisting of trastuzumab, bevacizumab, gemtuzumab, inotuzumab, polatuzumab, sacituzumab, adalimumab, infliximab, rituximab, and the combinations thereof.
In one embodiment of the invention, the active ingredient is a water-insoluble active ingredient, which is selected from the group consisting of vitamin A and its derivatives, Vitamin E and its derivatives, paclitaxel, docetaxol, camptothecin, doxorubicine, and curcumin.
In one example of the invention, the amphiphilic alginate has a molecular weight of 5,000 g/mole to 50,000 g/mole. The alginate salt is sodium alginate and has a molecular weight of 10,000 g/mole to 60,000 g/mole.
In one example of the invention, the hyaluronate is a hyaluronic salt and has a molecular weight of 100,000 g/mole to 1,000,000 g/mole, preferrably 100,000 g/mole to 500,000 g/mole.
In one example of the invention, the ionic crosslinker is selected from the group consisting of CaCl2), CaCO3, calcium phosphates, ZnCl2, BaCl2, and the mixture thereof. The gross concentration of the ionic crosslinker is from 0.5% to 5% (on gel weight base).
In one example of the present invention, the amphiphilic alginate nanoparticle is a fatty acid-conjugated alginate. The fatty acid-conjugated alginate is selected from the group consisting of oleic acid-conjugated alginate, stearic acid-conjugated alginate, linoleic acid-conjugated alginate, palmitic acid-conjugated alginate, and the combinations thereof. Preferably, the amphiphilic alginate nanoparticle is oleic acid-conjugated alginate.
According to the invention, the amphiphilic alginate-based nanoparticle can be used either alone or in combination with second drug being encapsulated in said amphiphilic alginate nanoparticle and allowing the composition to form a solid-like gel or high-viscous gel by crosslinking via the addition of metallic salts.
In another aspect, the present invention provides an injectable nanocomposite gel comprising an amphiphilic alginate-based nanoparticle and a salt of alginate and a hyaluronate, and a active ingredient and an ionic crosslinker or a mixture of the ionic crosslinkers.
In an embodiment of the invention, a low-molecular-weight alginate-based macromolecule is formed from an amphiphilic alginate or its derivatives (developed by Nuecology Biomedical Inc. Richmond, BC, Canada). According to the invention, the amphiphilic alginate is able to self-assemble into a nano-sized spherical nanoparticle in an aqueous environment which can be applicable to encapsulate hydrophobic In one specific example of the invention, amphiphilic alginate is a fatty-acid-conjugated alginate, and the active agent is a hydrophilic drug. T.
According to the invention, the said amphiphilic alginate nanoparticle can be used either alone or carries with a hydrophobic drug, further combining with gel matrix to ultimately develop a nanocomposite gel after gelation, where the final gel entity can be used for a subsequent injection to a subject for anti-tumor treatment. This fatty-acid-conjugated alginate nanoparticle exhibited excellent biocompatibility, drug loading ability and cellular uptake efficiency.
In a preferred embodiment of the present invention, the amphiphilic alginate can be used alone or in combination with an active ingredient, either water-soluble or water-insoluble, if practically needed, combined with highly porous gel matrix, to form a drug-carrying injectable nanocomposite gel. The porous gel matrix carried a water-soluble drug, which is used for specific anti-tumor treatment and the drug in the porous gel matrix can be a protein, an antibody drug, a biosimilar drug, an RNA-based molecule included but not limited to RNAi, microRNA, etc.
According to the invention, the porous gel matrix is composed of (1) a gel modifier, which included mid-to-high-molecular weight hyaluronate salts or its derivatives, (2) a gel former, which included low-molecular weight alginate salts in combination with low-molecular weight amphiphilic alginates, where the amphiphilic alginate is more preferable to have a cytotoxic potency to particularly cancerous cells or tissues, but is compatible to normal cells or tissues, (3) a gel stabilizer, included calcium chloride, (4) a gel crosslinker, which included but not limited to calcium chloride, calcium carbonate, barium chloride or zinc chloride, or metallic salts with divalent or trivalent coordination to those gel forming ingredients. The manufacturing procedures to produce resulting nanocomposite hydrogel are given below.
In another preferred embodiment, this invention provides the steps of:
According to the invention, the gel composition is used for drug delivery use. The said injectable gel was prepared by the method of the steps:
(i) preparing a mixture of alginate-based solution as Solution (1), where alginate salts and amphiphilic alginate with preferred weight ratio is prepared;
(ii) preparing a mixture of hyaluronates and metallic salts solution as Solution (2);
(iii) by mixing Solution (1) and Solution (2) with a ratio (by weight) from 0.5:1 to 2:1.
In one example of the invention, a high-viscous or solid-like gels can then be prepared by mixing Solution (1) with Solution (2), with gelation occurred in a manageable time period, to form a homogeneous nanocomposite gel. While adding biosimilar, antibody or protein drug, the drug was first dissolved and mixed in Solution (1) with a concentration ranging (in terms of final concentration in injectable gels) from 1.0% to 15% by weight, to form Solution (3). After then, by mixing Solution (2) and Solution (3), under continuous stirring, a final solid-like injectable gel can be formed for a subsequent medical uses.
The features and advantages of the present invention will be apparent to those skilled in the art. While numerous changes may be made by those skilled in the art, such changes are within the scope of this invention.
The foregoing summary, as well as the following detailed description of the invention, will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, there are shown in the drawings embodiments which are presently preferred. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities shown.
It is to be understood that, if any prior art publication is referred to herein, such reference does not constitute an admission that the publication forms a part of the common general knowledge in the art, in any countries or regions.
For the purpose of this specification, it will be clearly understood that the word “comprising or composed of” means “including but not limited to”, and that the word “comprises” has a corresponding meaning.
Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which the present invention belongs. One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. Indeed, the present invention is in no way limited to the methods and materials described.
According to the invention, an anibody or biosimlar or protein-like drug with high payload can be encapsulated by the said nanocomposite gel where drug potency can be enhanced to a large extent than that of free drug to against highly maligant tumor, take breast tumor as one exemplary case, under the same controlled protocol, and the drug-carrying injectable gel can be prepared in a specific and facile manner of production.
According to the invention, a vaccine with high payload can be encapsulated by the said nanocomposite gel where the vaccine efficacy can be enhanced to a large extent than that of vaccine alone to induce an immune response to recognize and fight against infective diseases, wherein the vaccine includes but not limited to whole pathogen vaccines, subunit vaccines, nucleic acid vaccines, and viral vectored vaccines.
Therefore, the present invention provides an antibody (or interchangeably, biosimilar as disclosed in this invention) drug-containing injectable gel, which includes a water-soluble active ingredient selected from the group comprising of trastuzumab, bevacizumab, gemtuzumab, inotuzumab, polatuzumab, sacituzumab, adalimumab, infliximab, and rituximab, a pharmaceutically acceptable biosimilar or interchangeably antibody drug derivative, either alone or in combination with a second water-insoluble active ingredient, comprising paclitaxel, docetaxel, doxorubicin, and curcumin, encapsulated in said amphiphilic alginate nanoparticle.
In some embodiments, the active ingredient is biosimilar drug or its derivatives.
According to the present invention, the amphiphilic alginate nanoparticles have hydrophobic and hydrophilic moieties to respectively interact with hydrophobic and hydrophilic molecules. The amphiphilic alginate carrier may include fatty-acid-conjugated alginate and/or derivatives thereof. Examples of said fatty-acid-conjugated alginate and derivatives thereof include, but are not limited to, oleic acid-conjugated alginate, stearic acid-conjugated alginate, linoleic acid-conjugated alginate, cholesterol-modified alginate. In an exemplary embodiment, the amphiphilic alginate-based nanoparticle is oleic acid-modified alginate.
According to the present invention, the antibody drug-containing injectable nanocomposite gel may use alone or further include an additional pharmaceutically active ingredient that is carried by the amphiphilic alginate nanoparticle. Examples of the additional active ingredient if pharmaceutically required, which is also water-insoluble includes, but are not limited to, Vitamin A and its derivatives, Vitamin E and its derivatives, anti-cancer drugs such as paclitaxel, docetaxol, camptothecin, doxorubicine, etc. According to the present invention, the said amphiphilic alginate nanoparticle has a particle size that ranges from 50 nm to 700 nm. In some embodiments, the said amphiphilic alginate nanoparticle has a particle size that ranges from 50 nm to 350 nm.
In addition, the present invention provides a method for anticancer drug in a subject, which includes administering to the subject the pharmaceutical composition by injection route described in this invention.
The pharmaceutical composition according to the present invention can be formulated into a dosage form suitable for injection administration using technology well known to those skilled in the art, which includes, but is not limited to, subcutaneous injection, intramuscular injection, intratumoral injection, and intraperitoneal injection.
The injectable nanocomposite gel according to this invention where the amphiphilic alginate nanoparticle plays a route not only capable of carrying a second pharmaceutically active ingredient if practically needed, which can be water-insoluble, but also acting as a buffer to accommodate the gelation rate of the injectable gel when the said Solution (2) and Solution (3) aforementioned were mixed. That is to say, the gelation time when those two solutions may become longer, from seconds to minutes or even prolonged, to ensure a final nanocomposite gel to be physically and chemically homogeneous for a subsequent use.
The invention will be further described by way of the following examples. However, it should be understood that the following examples are solely intended for the purpose of illustration and should not be construed as limiting the invention in practice.
1.1 Step 1
Solution (1) was prepared by mixing the gel stabilizer and/or crosslinker with structural modifier (hyaluronate salts which is employed to modify viscosity and homogenization of the resulting solution) into a first liquid medium.
1.2. Step 2
Solution (2) was prepared by mixing the amphiphilic alginates and alginate salts into a second liquid medium, which were acting as a dual-function ingredient for both gel former and drug carrier if practically required.
1.3. Step 3
Further mixing Solution (1) and Solution (2), stirring constantly, see below Drawing, to form a final nanocomposite hydrogel with a gelation time (from a viscous liquid to form a solid-like gel) ranging from 0.5 minutes to 20 minutes, depending on the concentration of CaCl2, CaCO3, ZnCl2, or BaCl2, as ionic crosslinker and the said gel former, i.e., amphiphilic alginate nanoparticles. The ionic crosslinker with a concentration of 0.5-5 wt % was used to form the injectable nanocomposite gel and the said amphiphilic alginate nanoparticle with a concentration of 0.05-2.0 wt %. The higher concentration of the amphiphilic alginate nanoparticle in said gel composition, the longer time period, for instance, from seconds to minutes or prolonged duration as increasing the amount of such amphiphilic nanoparticles, upon solid-gel development.
It is also important to learn the resulting injectable nanocomposite hydrogel can be prepared into a solid-like gel in both drug-free gel and trastuzumab-carrying gel (trastuzumab concentration is 10 wt % on weight base of the gel), where the gel viscosity is decreased significantly with increasing strain frequency, as shown in
The lower the gel viscosity under higher angular frequency is able to translate to a condition resemble that of syringe injection, which means the said nanocomposite gel and trastuzumab gel show shear-thinning behavior and allow to be injectable.
A shear-dependent storage modulus (G) and loss modulus (G′) is given in
The influence of ionic crosslinker concentration, taking CaCl2 or CaCO3 as one exemplary case, on the mechanical property of the nanocomposite gels without the presence of amphiphilic alginate nanoparticles, i.e., AGO2.0 composition, is given in
After the trastuzumab gel, with a drug concentration range of 2.5 wt %, 5 wt %, and 10 wt % (based on gel weight) was prepared, the drug-carrying gels were subjected to in-vitro drug release study,
Highly malignant breast cancerous SKBR3 cells were treated with Trastuzumab gel with drug concentration range of 0.5%, 1.0%, 2.5%, and 5%, respectively and respective controls, i.e., positive control and IgG negative control, as indicated in
The nanocomposite gel, with and without carrying T-mAb show a highly porous structure after freeze-dried as shown in
Acute toxicity of the drug-free injectable nanocomposite gel was carried out using ICR mice (n=10) for a time duration of 14 days. The gels with both AGO1.7 and AGO2.0 compositions were injected in an amount of 200 microliter each at subcutaneous site of the right flank region of the mice using a G30 syringe. The weight of the mice was monitored daily and remained constantly increase or similar during the test period. No measurable adverse effect was detected before sacrificed. Histopathological findings of the toxicity study for AGO1.7 and AGO2.0 compositions were examined, as illustrated in
The injectable nanocomposite gel carrying biosimilar drug, i.e., trastuzumab, with different dosing concentration designed based on clinical data per dosing, for a subsequent animal study. The breast tumor was cultivated by injection 1×107 SKBR3 cells to the right flank region of the mice, and the controls are given below:
It is encouraging that albeit the test is failed to effectively reduce or eliminate the tumor significantly for such a HER 2-positive highly-malignant breast tumor. Clinical standard to treat such HER 2-positive breast tumor is using trastuzumab drug or Herceptin®, via SC injection or vein injection, this invention disclosed a new opportunity to use trastuzumab gel where an enhanced therapeutic performance in inhibiting the growth of SKBR-3-derived tumor can be clearly observed, improved by a factor of 2-3 times the size change during the test period, comparing to both control group and free-trastuzumab group. This suggests the use of trastuzumab gel disclosed in this invention improved therapeutic efficacy to a considerable extent, and is worthy of moving toward a potential clinical translation for further application.
Two AGO-based nanocomposite injectable gels were prepared as illustrated in
In-vitro cell viability test was carried out using free paclitaxel and PTX-T-mAb gel over a cell culture condition given as:
The resulting cell viability is given in
The injectable nanocomposite gel carrying both chemo-drug, i.e., paclitaxel, and biosimilar drug, i.e., trastuzumab (T-mAb), with different dosing concentrations designed based on clinical data per dosing, for a subsequent animal study. The breast tumor was cultivated by injection 1×107 SKBR3 cells to the right flank region of the mice, and the controls are given below:
The resulting tumor size measurement over the time duration (15-day duration) of animal study is illustrated in
This study also indicates a sustain release of both water-insoluble chemo-drug, paclitaxel, and water-soluble, antibody drug T-mAb, that can be co-delivered effectively against highly-metastasized HER2-positive breast tumor with synergy, compared with co-administration of both drugs in their free form.
It is encouraging that a co-delivery and co-release of anti-breast tumor drugs of distinct physico-chemical and therapeutic properties through the use of injectable AGO-based gel for SC administration can be technically and therapeutically achieved in the prevention of metastasized HER2-positive breast tumor from substantiated growth in animal body.
Clinical standard to treat such HER 2-positive breast tumor is typically using trastuzumab drug or Herceptin®, or a combination therapy of T-mAb and chemo-drug administrated in sequential manner via mostly vein injection or some via SC injection, this invention disclosed a new opportunity to use AGO-based injectable gel to carry a single high-dose Biosimilar drug or a combination of Biosimilar drug, i.e., T-mAb, and a traditional chemo-drug, i.e., paclitaxel, followed by co-releasing both drugs from the gel where an enhanced therapeutic performance in inhibiting the growth of SKBR-3-derived tumor, as model tumor, can be clearly observed, improved by a factor of 2-4 times the tumor size change during the test period, comparing to both control group and free-T-mAb group. This suggests the use of AGO-based nanocomposite gel disclosed in this invention improved therapeutic efficacy to a considerable extent, and is worthy of moving toward a potential clinical translation for further anti-cancer application.
All patents and references cited in this specification are incorporated herein in their entirety as reference. Where there is conflict, the descriptions in this case, including the definitions, shall prevail.
While the invention has been described in connection with what are considered the exemplary embodiments, it is understood that this invention is not limited to the disclosed embodiments but is intended to cover various arrangements included within the spirit and scope of the broadest interpretation so as to encompass all such modifications and equivalent arrangements.
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CA2021/050955 | 7/12/2021 | WO |
| Number | Date | Country | |
|---|---|---|---|
| 63050396 | Jul 2020 | US |
