KR20200042440A - Parenteral non-systemic administration of buffers to inhibit solid tumors, hyperpigmentation and metastasis of gout - Google Patents

Abstract

Parenteral administration (except for direct systemic administration) of the present invention, alone or in combination with a pH-increasing agent or other metastasis inhibitor, is effective and overcomes the disadvantages of intravenous and oral administration in the treatment of solid tumors, scleroderma and gout.

Description

Parenteral non-systemic administration of buffers to suppress metastasis, hyperpigmentation and gout of solid tumors

Cross-reference to related applications

This application is filed on U.S. Provisional Application No. 62 / 602,227 filed on April 17, 2017, U.S. Provisional Application No. 62 / 602,358 filed on April 20, 2017, and U.S. Provisional Application No. 62 filed on September 18, 2017 / 559,947, US Provisional Application No. 62 / 559,360 filed on September 15, 2017, US Provisional Application No. 62 / 562,725 filed on September 25, 2017, US Provisional Application No. filed on September 22, 2017 Priority is claimed on 62 / 609,982, and U.S. Provisional Application No. 62 / 639,904, filed on March 7, 2018. The contents of these patent documents are incorporated herein by reference in their entirety.

Technical field

The present invention relates to the use of anti-metastatic agents, including buffering agents, which are administered parenterally, non-systemically, particularly topically to create resistance to metastasis of the tumor. Buffers that increase pH are also useful in treating hyperpigmentation and gout. It is believed that both non-systemic topical and other parenteral administrations themselves overcome the difficulties of oral administration and systemic parenteral administration of buffers designed to raise local pH.

Progression to metastasis remains the highest risk of death for cancer patients despite significant efforts to therapeutically target metastatic lesions. Tumor invasion and metastasis associated with tumor progression are the leading cause of cancer death, and understanding the mechanisms that determine the spread of metastatic spread of malignant cells through invasion into distal tissue is probably an important problem in oncology.

Microenvironment acidosis in primary tumors increases cell mobility and invasiveness, thereby increasing metastasis, and solid tumors have a relatively low pH, probably due to the hypoxic state of these tumors, increased glycolysis metabolism of glucose, and insufficient perfusion. It is known to exist in the microenvironment. In 1979, Turner, GA, Experientia (1979) 35: 1657-1658 reported that acid pH promotes metastasis by promoting the release of tumor cells by collagenase. Next literature [Curvier, C. et al. , Clin. Exp Metastasis (1997) 15: 19-25] reported that glucose starvation, hypoxia and acidosis improved tumor cell invasion. Next article [Rofstad, EK et al., Cancer Res (2006) 66: 6699-6706] reported that the acidic extracellular pH of human melanoma cells promotes metastasis in mice.

The extracellular pH of malignant solid tumors is acidic in the range of 6.5 to 6.9, but the pH of normal tissues is significantly more alkaline of 7.2 to 7.5. These observations led to the “acid-mediated invasion hypothesis,” in which the tumor-derived acid promotes tumor invasion by promoting normal cell death and extracellular matrix (ECM) degradation of the parenchyma surrounding the growing tumor. Promote.

According to various sources in the public newspaper, certain doctors have been experimenting with intravenous sodium bicarbonate as a way to suppress metastatic cancer.

A series of articles by Robey, IF et al., Starting with a publication in Cancer Res (2009) 69: 2260-2267, demonstrated that oral bicarbonate reduced the formation of spontaneous metastasis in a mouse model of metastatic breast cancer. See Lobey, IF et al., BMC Cancer (2011) 11: 235-245 and Robey, IF, et al. BioMed Res International (2013) pages 1-10 and Robey, IF et al . , J. Integr. Uncol . (2015) 4: 1-8] described oral administration of bicarbonate as a metastasis inhibitor in mice and subsequent human volunteers.

The following literature (Ribeiro, M. de L., et al., J. Nutr Food Sci (2013) 2: 6-16) gave foods (including wine) various pH scores, and lysine buffers in mice with prostate cancer and Oral administration of bicarbonate was described. In addition, the following document (Silva, AS et al ., Cancer Res (2009) 2677-2684) described the role of systemic buffers in reducing metastasis. Sircus, M., published a book, Sodium Bicarbonate: Nature's Unique First Aid Remedy, Garden City Park, New York: Square One Publishers, 2014, advocating sodium bicarbonate as a treatment for various conditions.

However, oral administration of buffers has been found to have side effects including diarrhea, nausea, vomiting and abdominal discomfort. In addition, intravenous administration, that is, systemic administration, has also been distrusted.

Treatment of cancer and prevention of metastasis are important applications of the methods of the present invention, but systemic regulation of pH is also important in other situations. In this context, for example, treatment of pain is included. In addition, in recent unpublished results, it has been demonstrated by the ET Science of South Pasadena in California that topical application of sodium bicarbonate is effective in reducing blood lactate after exercise and improving performance in cycling.

However, not all cancers respond to the adjustment of the microenvironmental pH by administration of buffers. The following literature (Bailey KM et al., Neoplasia , (2014) 16: 354-364) showed that metastasis was not inhibited by the buffer in some tumor models regardless of the buffer used. In cell phenotypes that utilize a pH-dependent mechanism for successful metastasis, high glycolysis phenotypes acidify the local tumor microenvironment resulting in morphological changes, whereas buffer resistant cell lines do not undergo matrix-degrading protease ( matrix-degrading protease).

In addition to simple buffering to adjust pH, undesirable extracellular pH problems can be solved using alternative target DNA ⁺ / H ⁺ exchanger 1 (NHE1) proteins, which may be targets of transfer inhibitors. For example, Loo, SY et al., Curr. Pharm. Des. (2012) 18: 1372-1382 describes a promising drug target NHE1 as an anti-cancer strategy. Therefore, therapeutic agents targeting these proteins are also useful in the present invention. [Amith, SR and Fliegel, L. Cancer Res. (2013) 73: 1-6] lists a number of factors that make up a suitable target by affecting these proteins.

A number of other factors are also associated with tumor metastasis, and in many cases, therapeutic agents targeting them are actually available. Among these are matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) and certain cathepsins, especially cathepsins B, D and L. For example, cathepsin B as a cancer target is described in Gondi, CS and Rao, JS Expert Opin. Ther. Targets (2013) 17: 281-291, and cathepsins are generally targeted in the following literature [Olson, OC and Joyce, JA, Nat. Rev. Cancer (2015) 15: 712-729. For a general discussion of the role of MMPs in the progression of cancer, see Gialeli, C. et al ., FEBS J. (2011) 278: 16-27. According to this article, MMP plays a broad role in modeling extracellular matrix, allowing cancer cells to escape from solid tumors.

Small molecule inhibitors of these proteins are known, for example, various N-sulfonyl amino acid derivatives are described in Tamura, Y. et al ., J. Med. Chem . (1988) 41: 640-649, and since then a number of additional agents have been developed. In addition, naturally occurring endogenous cysteine protease inhibitors are present. Endogenous CPIs include a single protein superfamily of cystatins, such as type 1 cystatin, stepin A (or cystatin A) with a molecular mass of 11,775 Da and stepin B (or cystatin B) with a molecular mass of 11,006 Da. Make up. Cysteine protease inhibitors (CPIs) are pseudo-irreversible inhibitors that bind very tightly.

Another transfer inhibitor is an inhibitor of the src homology region 2-containing protein tyrosinase phosphatase (Shp2). Many of these activities, including Fumosorine, PHPS (NSC-87877) and NSC-117199, phenylhydrazonopyrazolone sulfonate (PHPS1), DCA, cryptotanshinone, 11-B08 and # 220-324 Inhibitors are known.

Another metastasis inhibitor is based on the well-known role of epidermal growth factor receptor (EGFR) and / or integrin in inducing tumor progression, and it is well known that all of these classes of receptors promote NHE1 activity. Several anti-EGFR compounds (eg, erlotinib) have been approved to inhibit metastasis, and anti-integrin drugs (cilengitide) are in clinical trials. Cariporide, eniporide and / or amiloride have passed all clinical stages.

Finally, phytochemicals with antioxidant and anti-inflammatory properties are thought to play an important role in the prevention and / or treatment of cancer. So far, various anti-clinical experimental models have been used to investigate the potential anti-cancer properties of numerous phytochemicals.

In particular, a number of laboratory-based studies have demonstrated the anti-cancer effect of the medicinal plant, Withania somnifera . These anti-cancer properties are due to the withanolides, a class of bioactive components that include Wittaferin A (WFA). Witanolide is a group of naturally occurring C28 steroid lactones. These include 4 cycloalkane ring structures, 3 cyclohexane rings and 1 cyclopentane ring. WFA is a steroidal lactone derived from Acnistus arboescens , Withania somnifera and other members of the Solanaceae family . It has been traditionally used in ayurvedic medicine. The first member of the witanolide class of the ergostan type product was discovered.

Many of the aforementioned agents can be used in combination as described below.

Parenteral non-systemic administration of these buffers and agents, including topical administration of biocompatible buffers and other metastasis inhibitors, has a positive effect on cancer, including inhibition of metastasis, inhibition of growth, and in some cases tumor contraction. It has been confirmed to affect. In addition, such administration of a pH-enhancing buffer is useful for treating gout and dermatosis (hyperpigmentation).

Topical administration can utilize a penetrant that improves transdermal passage topically or systemically. Suitable penetrants are described, for example, in PCT publications WO / 2016/105499 and WO / 2017/127834.

Various substances, including tyrosinase inhibitors hydroquinone, kojic acid, azelaic acid, various phenols, and arbutin, have been proposed for the treatment of dermatosis. They appear to be completely unsuccessful and have undesirable side effects. The causes of dermatosis are diverse and their condition is characterized by a number of hyperpigmented lesions of varying sizes, shapes and distributions. These lesions can be appropriately treated by raising the local pH according to the methods of the invention described below.

The contents of all documents cited above, such as the contents of any cited document throughout this specification, are incorporated herein by reference.

Disclosure of the invention

Applicants have confirmed that the disadvantages of intravenous and oral administration of buffers and other metastasis inhibitors can be overcome by non-systemic parenteral administration, such as intraperitoneal, subcutaneous or intramuscular injection, particularly topical administration. . Topical administration is the most convenient transdermal administration, but further includes transmembrane administration, for example by suppository or intranasal application.

Thus, in one aspect, the present invention is a method for inhibiting metastasis and / or growth of a solid tumor contained in a subject, and an effective amount of an anti-metastasis agent is administered to a subject in need of such inhibition. It relates to a method comprising administering by a non-systemic transdermal route. In some embodiments, the metastasis inhibitor is a buffer or protease inhibitor sufficient to increase the pH of the microenvironment of a solid tumor, an inhibitor of Na ⁺ / H ⁺ exchange activity, an inhibitor of epidermal growth factor receptor (EGFR), src homology region 2 Inhibitors of the containing protein tyrosinase phosphatase (Shp2), withaferin A, or a combination thereof. The administration is via a non-systemic parenteral route and includes topical administration, particularly transdermal administration.

Particularly for transdermal topical administration of agents other than buffers, suitable formulations generally include a penetrant that enhances skin penetration, and in some embodiments are composed of chemical penetration enhancers (CPEs). In some cases it may also include a peptide designed to penetrate cells, i.e. a cell penetrating peptide (CPP), also known as a skin penetrating peptide (SPP). The formulation may be applied as such or by an occlusive device such as a patch.

When the active agent is a buffer, the selection of the buffer system is based on the biocompatibility of the buffer system itself and the compatibility of the buffer system with other components of the formulation, as well as the criteria for buffer capacity at a suitable pH, typically between 7 and 8. Conversely, the agent is selected to be compatible with the selected buffer; The amount of penetrant is generally less than the amount beneficial to the therapeutic agent.

When the pH is adjusted for the purpose of inhibiting metastasis, an efficacy assessment is performed after treatment. More importantly, as mentioned above, some tumors have been found to be resistant to buffer treatment, ie, raising the pH has no metastasis inhibitory effect. For example, Bailey, KM, et al. , Neoplasia (2014) 16: 354-364 (mentioned above). Thus, it is one aspect of the present invention to evaluate the tumors of potential subjects for treatment with buffers, for example by culturing biopsy for sensitivity / resistance to pH control.

Elevating pH in the vicinity of black dermatosis and gout is also an effective treatment. The method of the present invention can also be usefully applied to the treatment of this condition. Accordingly, another aspect of the present invention relates to this method of treatment for obesity and gout.

1 shows the time course of urine pH immediately after topical administration of sodium bicarbonate with various formulations and dosing regimens.
FIG. 2 shows the average daily urine pH of a test subject over time after administration of a formulation of sodium bicarbonate with the formulation and dosage of FIG. 1.
3 shows the time course of urine pH over a period of 3 days using an alternative topical formulation.

Mode of carrying out the invention

As noted above, one aspect of the present invention includes the administration of non-systemic parenteral administration of cancer mass by topical administration of a metastasis inhibitor comprising an agent that buffers the immediate environment of tumor cells, including solid tumors and melanoma. It is a method of inhibiting cancer growth and metastasis, including reduction. For non-systemic parenteral administration, such as intramuscular, intraperitoneal or subcutaneous administration, a standard formulation is sufficient. These formulations include standard excipients and other auxiliary ingredients such as antioxidants, suitable salt concentrations, and the like. Such formulations can be found, for example, in the latest edition of Remington's Pharmaceutical Sciences , Mack Publishing Company, Easton, PA, the standard reference for various types of administration.

Since the subject of the method of the invention is a veterinary subject in addition to humans, preparations suitable for these subjects are also suitable. Such subjects include domestic animals and pets, as well as sports animals such as horses and greyhounds.

One important aspect of the invention is that some tumors are buffered because the microenvironment of the tumor is not acidic and at least some of these tumors achieve metastasis by elevation of specific proteolytic enzymes that degrade extracellular matrix (ECM). It is based on the above-mentioned perception that it does not respond to. When buffer treatment is considered, it is preferred that tumor cells from a biopsy of a solid tumor in a subject are cultured and tested prior to treatment to ensure responsiveness to the buffer. These evaluations were conducted in Bailey, K.M. As described in et al. (2014), it can be performed by any suitable means, including pH measurement, evaluation of the level of the relevant protease, and invasion analysis affected by buffering. One important such analysis is the glycolysis stress analysis as described herein. Cell cultures of biopsy tumors that do not appear to respond to buffer treatment, as confirmed by this analysis, may be useful for administration of other metastasis inhibitors and it may be beneficial to include these agents in compositions of the invention comprising buffers. have. Therefore, treatment with the buffer-containing composition alone may be contraindicated, and the buffer is not administered as the only active agent in the subject, but is useful for alternative treatments. Needless to say, this does not necessarily mean that the buffer is omitted from the formulation used to administer the alternative active agent.

For topical administration, particularly transdermal administration, the agent comprises one or both of chemical penetrants (CPE) and peptidic cell penetrants (CPP) that promote delivery across and / or across the dermis, including membranes. Penetrating agents may be included, especially in the case of transdermal administration as well as suppository or intranasal administration. Particularly suitable penetrants for those containing at least one agent other than a buffer are those described in WO2016 / 105499 and WO2017 / 127834 mentioned above. In addition to formulations containing penetrants, transdermal delivery can be achieved by mechanically destroying the skin surface to promote penetration or simply by supplying formulations applied to the skin under an occlusive patch.

Briefly, the penetrants described in the aforementioned PCT applications are based on a combination of synergistically acting components. Many such penetrants are used in combination with a lecithin organogel present in the penetrant to provide 25 to 70% by weight of the formulation, and an alcohol such as benzyl alcohol, which provides a concentration of 0.5 to 20% by weight of the final formulation. It is based. In general, such formulations are essentially polar formulations, for example aqueous formulations, or alternatively additional ingredients, comprising a nonionic surfactant sufficient to be present at 1-25% by weight of the final formulation. An anhydrous formulation comprising a bile salt sufficient to provide 1-15% by weight of the formulation and a surfactant present to provide 20-60% by weight of the formulation. In addition, these penetrants are useful when the transfer inhibitor is a buffer, but a lesser amount of lecithin organogel may be required, for example, 1 to 50% by weight of a lecithin organogel if buffer is present at high concentration. You can. Lecithin is exemplified, but other amphoteric temperatures can be used.

Alternatively or additionally, the penetrant may provide one or more electrolytes sufficient to impart viscosity and viscoelasticity, as well as an equimolar mixture of approximately 1: 1: 1 of the bile salt: lecithin and completion component. It is a degree to include at least a surfactant and benzyl alcohol or analogs thereof. The finished component can be a polar liquid, a non-polar liquid or both amphiphilic substances, but both amphiphilic substances are very preferred.

In particular, details regarding the behavior of this type of composition in aqueous solvents are described in Chen, CY, the disclosure of which is hereby specifically incorporated by reference herein for a method of preparing an advantageous micellular composition for transdermal administration of the active agent. , et al, Langmuir (2014) 30: 10224-10230. As described, the mixture of lecithin and bile salts in an approximate 1: 1 molar ratio, particularly in the presence of an electrolyte, forms a stable sticky wormlike colloid in water. These colloids help to deliver active agents, especially high molecular weight agents, across the skin. Although a molar ratio of 1: 1 is optimal, Applicants have confirmed that modifications from this ideal can be used. That is, when the finished component of the "1: 1: 1" system is a fatty acid such as isopropyl palmitate, a molar ratio of 1: 0.5 to 1: 5 may be used. Thus, “approximately 1: 1: 1 molar ratio” refers to this range.

In any case, the penetrant is a keratinase and / or skin permeation peptide (sometimes called a skin permeation peptide) and / or a penetration enhancer effective to reduce thiol bonds, break hydrogen bonds, and / or affect keratin dissolution. (permeation enhancer) may be further included. The additional ingredients listed above are 1: 1 to an equimolar ratio of salts: lecithins: complete to penetrants and formulations that provide 25 to 70% by weight of lecithin organogel and 0.5 to 20% by weight of benzyl alcohol by weight of the final formulation. It may be included in all of the above penetrants comprising sufficient salt: lecithin: finished ingredients to provide the ingredients.

As an alternative to topical administration to intact skin, the skin surface is mechanically destroyed by the use of spring systems, laser driven systems, systems driven by the Lorentz force, or by shock waves including gas or ultrasound. And can be destroyed, for example, using microdermabrasion using sandpaper or equivalents, or using a microneedle or electroporation device. A simple solution of the agent (s) as well as the agents listed above that penetrate the intact skin can be applied using a closed patch, such as in the form of a micro patch. In addition, external reservoirs of the formulation for extended administration may be used.

The active buffer component is any weakly basic compound or combination that produces pH 7-8 in the microenvironment of tumor cells. Such buffers in addition to or instead of bicarbonates include lysine buffers, chloroacetate buffers, tris buffers (i.e., buffers using tris (hydroxymethyl) aminoethane), phosphate buffers and non-natural amino acids with similar pKa values as lysine. There are buffers that use. They can be, for example, enantiomers of the natural form of these amino acids, or lysine analogs with long or short carbon chains or branched forms thereof. In addition, histidine buffers can also be used. Generally, the concentration of buffer in the composition ranges from 1 to 40% wt / wt. The more common range for sodium bicarbonate or sodium carbonate or both as a pH adjusting agent is 5 to 35% by weight. The absence of inflammatory side effects at these concentrations is surprising because the use of lower concentrations to treat other conditions such as arthritis is mentioned in US 5,176,918.

In all of the above cases, additional anti-cancer ingredients may be used, such as chemotherapy agents such as irinotecan, floxuridine, daunorubicin, cytarabine, and the like.

As mentioned above, some tumors do not respond to treatment with buffers, but are understood to metastasize due to elevated levels of protease attacking the extracellular matrix surrounding the tumor. In any event, degradation of ECM will promote metastasis. Accordingly, additional active agents optionally included in the compositions of the present invention are one or more suitable protease inhibitors. Of particular interest are cathepsin inhibitors, such as inhibitors of cathepsin B and substrate protease (MMP). These components alone enhance the effectiveness of the buffer on tumors that are active or not resistant to buffer treatment.

Another active agent is Wittaferrin A. Wittaferrin A inhibits tumor metastasis and exhibits other anti-cancer activity, such as inhibition of neovascularization and cell proliferation associated with carcinoma. In addition, Witaperin A is a leptin sensitizer with strong anti-diabetic properties that can induce beneficial effects on healthy weight loss and glucose metabolism.

In addition to simple buffering to adjust pH, undesirable extracellular pH problems can be solved using alternative target DNA ⁺ / H ⁺ exchange 1 (NHE1), which may be the target of metastasis inhibitors. Another transfer inhibitor is an inhibitor of the src homology region 2-containing protein tyrosinase phosphatase (Shp2). A number of inhibitors of this activity are known, including Fumosorine, PHPS (NSC-87877) and NSC-117199, phenylhydrazonopyrazolone sulfonate (PHPS1), DCA, cryptotansinone, 11-B08 and # 220-. 324, metalloproteinase-2 and -9 (MMP-2 and MMP-9) and certain cathepsins, especially cathepsins B, D and L.

Other agents include inhibitors of E-cadherin and inhibitors of epidermal growth factor receptor (EGFR). Known inhibitors include erlotinib, anti-integrin drugs (silengitide), caripoliride, enipolide and amiloride.

The formulation may contain other ingredients that act as excipients or serve purposes other than the active anti-tumor effect. For example, antioxidants such as preservatives and antibacterial agents such as ascorbic acid or α-lipoic acid can be included. Ingredients other than therapeutically active ingredients and ingredients that are the main effector of skin penetration include ingredients provided for aesthetic purposes, such as menthol or other aromatics, and ingredients that affect the physical state of the composition, such as emulsifiers and emulsifiers. For example, Durasoft (this is a mixture of thermoplastic polyurethane and polycarbonate). Generally, these ingredients are present in the composition in very small proportions. It is understood that these latter adjuvants are not therapeutic ingredients and are not primarily responsible for skin penetration. Ingredients that primarily affect skin penetration have been described in detail as described above. However, some of these substances have some ability to affect skin penetration. For example, the following document describing the penetration properties of menthol [Kunta, JR et al ., J. Pharm. Sci. (1997) 86: 1369-1373. That is, their presence in the formulations useful in the present invention is not necessary to achieve transdermal transport of the active agent (s), and they can be omitted from the formulation.

In some embodiments, the final formulation for transdermal administration may be in the form of a colloidal particle composition. Typical colloidal particle dimensions range from 10 to 100 nm. Methods of providing colloidal morphology are known in the art. The formation of colloidal particles is facilitated not only by mechanical agitation, but also by an appropriate level of components capable of forming colloidal particles. (See Cheng, C-Y et al. (2014) mentioned above). Various devices are commercially available to promote the formation of colloid particles.

For example, the formation of colloidal particles is enhanced by milling. The level of improvement is determined not only by the number of passes through the mill, but also by the pressure and speed at which the milling occurs. As the number of passes and the speed and pressure increase, the level of colloid formation is improved.

If the grinder is Dermamill 100 sold by Medisca® (Blaubrite), the typical speed includes random fluctuations between 1 and 100, where 1 is the slowest and 100 is the fastest. The pressure is selected from 1 to 5, where 1 is the highest and 5 is the lowest. In some embodiments, multiple passes are considered, such as 2 to 10 or more passes. Median values are also included. The speed and pressure can be changed for each pass.

Alternative grinders can also be used, and the speed, pressure and number of passes can be replicated similarly compared to Dermamill 100 equivalents. Alternatively, colloidal particle density can be compared microscopically to ensure results equivalent to those obtained with Dermamill 100. In some embodiments, the colloidal particle density is at least 20%, and in many cases at least 30%.

Generally, the size of colloid particles is in the nanometer range, for example 10 to 150 nm, including the median value.

Formulations of this type are particularly important for formulations containing bile salts. In an embodiment in which bile salt is added to the combination of benzyl alcohol and lecithin organogel instead of topping with an aqueous medium, the relatively spherical colloid particles can elongate into the shape of a long worm, thus allowing excellent penetration of the epidermal stratum corneum. The worm-like formation of the colloidal particles is particularly helpful in accommodating high molecular weight therapeutic agents. As is known, bile salts are amphiphilic substances on the face and include salts of taurocholic acid, glycocholic acid, taurochenodeoxycholic acid, glycokenodeoxycholic acid, cholic acid, and deoxycholic acid. In addition, detergents are useful instead of bile salts and include Tween 80 and Span 80.

Thus, the inclusion of bile salts promotes ultraformability of colloidal particles that promote the passage of low and high molecular weight drugs and other active agents, such as nucleic acids and proteins. These compositions overcome the skin penetration barrier by squeezing along the natural gradient across the stratum corneum by squeezing itself along the intercellular sealing lipid. This promotes local and reversible changes in the membrane composition when compressed or pulled into narrow pores.

Thus, bile salts in combination with lecithin organogels promote factors of colloidal particle stability, improved viscosity and viscoelasticity, which are important for transdermal drug delivery. Both thermodynamic and kinetic stability are improved by the addition of background electrolytes such as sodium chloride and sodium citrate. These electrolytes can increase the viscosity and viscoelasticity of colloidal particles more effectively and block repulsion between bile salt anions at minimal concentrations.

In some embodiments, the molar ratio of bile salt to lecithin is 1: 1, and the concentration of the electrolyte is determined by titration of the solution for improved viscosity and clarity as measured when the container of the solution is turned over.

Formulations designed for transdermal administration of the active ingredients, particularly buffers and other therapeutically active compounds and any auxiliary ingredients, can be tested with a variety of known protocols for their properties and their effect on achieving passage across the skin. have. For example, the level of colloid formation can be determined by rheological studies using a commercially available flow meter, the RDA-3 strain control rheometer. The level of collagen and proteoglycan components of the extracellular matrix in the skin from subjects exposed to the penetrating composition can be measured histologically using, for example, H & E and Alitian Blue. The skin model was prepared as a three-dimensional structure using human-derived epidermal keratinocytes and cultured in normal human-derived dermal fibroblasts to create a multi-layer, highly differentiated model of human dermis and epidermis. In addition, moisture loss measurements may also be appropriate and these measurements may be measured using a commercially available dermalab evaporimetry system commercially available from Cortex Technology, Hadsun, Denmark. In addition, collagen message levels can be measured using standard techniques, and the permeability of human epidermis can be measured by determining electrical conductivity. Monitoring the electrical conductivity of the skin exposed to various formulations can identify the most efficient agents for increasing permeability. In addition, by using trace elements such as copper and iron, it is possible to confirm the penetration effect, such as chromatographic analysis of components passing through the skin layer.

For illustrative purposes only, the following are examples of integrated cooperative CPE preparations for extracellular matrix.

-Cetyltrimethyl ammonium bromide (about 2.0% to about 10.0%)

Sodium cholate: lecithin (96% purity): isopropyl myristate (equivalent to 1: 1: 1, about 10% to about 40.0%)

-Sodium citrate (Titration with # 2 transparency / increased viscosity)

-Benzyl alcohol (about 2.0% to about 30.0%)

-Cis-palitoleic acid (about 20.0% to about 30% BA)

-Methyl pyrrolidone (0.4%) / dodecyl pyridinium (1.1%) (about 0.5% to about 5.0%)

-Pluronic 127 (Amount that makes 100%)

An example of a CPP agent for cellular components of the SC permeation barrier is as follows:

ACSSSPSKHCG identified by TD-1 [alanine-cysteine-serine-serine-serine-proline-serine-lysine-histidine-cysteine-glycine]

-Thioglycolic acid (TGA) (about 2.0% to about 7.0% concentration) (can be replaced by other reducing agents)

Proteinase K (about 5 mg / mL to about 15 mg / mL)

Aspects of the invention, including administering a buffer to locally raise the pH in the environment of a solid tumor or in the vicinity of gout or scleroderma, extend the indications to which the methods of the invention can be applied. In general, for buffers as active agents, the requirements for effective penetration of the skin have been found to be less restrictive than those required for alternative agents useful in preventing cancer metastasis. Although the penetrants detailed above are useful, for example, a wider range of concentrations of lecithin organogels are effective in this case. In some cases, in addition to lecithin in organogels, additional lecithin is included. In addition, for such indications, delivery of solid tumors including melanoma to melanoma or gout to the locus is preferred, but effective systemic pH changes can be used as a method of diagnosing the effect of infiltration when topical administration is used. Thus, in some of the following examples, an increase in the pH of urine in a model system such as a mouse can be used to verify the effectiveness of a penetrant in a formulation, but the effect on pH is local to tumors, dermatosis or gout. desirable.

The following examples are intended to illustrate but not limit the invention.

Example 1

Certain formulations for the administration of buffers

The following composition was prepared and found to be useful in the method of the present invention. In the tables below, “LIP” refers to a lecithin organogel consisting of a 1: 1 molar mixture of soybean lecithin and isopropyl palmitate containing 96% phosphatidyl choline; "BA" represents benzyl alcohol; PLU-F127 refers to the detergent Poloxamer F127 granules; PLU-water represents PLU-F127 dissolved in deionized water. (Alternatively, a commercially available Pluronic F127 30% gel can be used) Durasoft® is an emulsifier in a commercially available form.

In the tables below, sodium bicarbonate and sodium carbonate were fed as-is. Tris buffer of pH 8.1 was used. Phosphate “buffer” was supplied as NaH ₂ PO ₄ . Percentages are weights per weight (wt / wt), ie weight percentages.

Bicarbonate preparation IB IIB LIP 30.00% 25.00% ethanol 1.50% 0.00% BA 1.00% 1.00% menthol 0.50% 0.90% Sodium bicarbonate 33.50% 5.76% PLU-F127 10.05% 10.50% Water in gel with PLU 23.45% 24.50% Additional water 30.34% Durasoft 2.00% Sum 100.00% 100.00%

Carbonate preparations IC IIC IIIC IVC LIP 25.00% 25.00% 30.00% 40.00% ethanol 0.00% 0.00% 1.50% 1.50% BA 1.00% 1.00% 1.00% 1.00% menthol 0.90% 0.90% 0.50% 0.50% Sodium carbonate 9.36% 16.10% 33.50% 11.00% PLU-F127 10.50% 10.00% 10.05% 13.50% Water in gel with PLU 24.50% 23.33% 23.45% 31.53% Additional water 26.74% 22.67% 0.00% 0.00% Durasoft 2.00% 1.00% 0.00% 1.00% Sum 100.00% 100.00% 100.00% 100.00%

Tris formulation IT IIT IIIT IVT LIP 25.00% 25.00% 30.00% 38.40% ethanol 0.00% 0.00% 1.50% 1.50% BA 1.00% 1.00% 1.00% 1.00% menthol 0.90% 0.90% 0.50% 0.50% Tris buffer, pH 8.1 13.37% 23.00% 33.50% 14.93% PLU-F127 10.50% 10.00% 10.05% 12.80% Water in gel with PLU 24.50% 23.33% 23.45% 29.87% Additional water 22.73% 15.77% 0.00% 0.00% Durasoft 2.00% 1.00% 0.00% 1.00% Sum 100.00% 100.00% 100.00% 100.00%

Phosphate preparation IP IIP IIIP IVP LIP 25.00% 25.00% 30.00% 37.25% ethanol 0.00% 0.00% 1.50% 1.50% BA 1.00% 1.00% 1.00% 1.00% menthol 0.90% 0.90% 0.50% 0.50% Sodium phosphate 16.02% 27.55% 33.50% 17.36% PLU-F127 10.50% 10.00% 10.05% 12.42% Water in gel with PLU 24.50% 23.33% 23.45% 28.97% Additional water 20.08% 11.22% 0.00% 0.00% Durasoft 2.00% 1.00% 0.00% 1.00% Sum 100.00% 100.00% 100.00% 100.00%

Carbonate, tris, and phosphate combination preparations Phosphate-low Phosphate-high III IV LIP 25.00% 25.00% 30.00% 37.25% ethanol 0.00% 0.00% 1.50% 1.50% BA 1.00% 1.00% 1.00% 1.00% menthol 0.90% 0.90% 0.50% 0.50% Sodium carbonate 5.30% 5.30% 5.30% 5.30% Tris (121.14) 6.00% 6.00% 6.00% 6.00% Sodium phosphate 6.00% 6.00% 6.00% 6.00% PLU-F127 10.50% 10.00% 10.05% 12.42% Water in gel with PLU 24.50% 23.33% 23.45% 28.97% Additional water 12.14% 21.47% 16.20% 0.06% DURASOFT 2.00% 1.00% 0.00% 1.00% Sum 100.00% 100.00% 100.00% 100.00%

Alternative buffer 25 28 29 A (2) B (2) C (2) LIP 6.00% 12.00% 12.00% 14.00% 15.00% 18.00% BA 1.00% 1.00% 1.00% 1.00% 1.00% 1.00% menthol 0.50% 0.50% 0.50% 0.25% 0.25% 0.50% Durasoft 1.50% 1.50% 1.00% Pluronic granules 4.20% 4.20% 4.20% 5.40% 2.10% 3.60% water 37.80% 37.80% 40.30% 31.60% 29.65% 41.90% Sodium carbonate 7.00% 7.00% 7.00% *? *

3.00% Sodium bicarbonate 28.00% 28.00% 28.00% 32.50% 32.50% 15.00% Propylene glycol 3.00% 3.00% 6.00% 10.00% 5.00% Almond oil 3.00% 3.00% 3.00% 4.00% 3.00% 4.50% Zinc oxide 0.25% 0.50% Cetyl alcohol 2.00% 2.00% 2.00% 2.00% 3.00% 3.00% lecithin 3.00% Cetiol ultimate (a mixture of tridecane and undecane) 3.00% ethanol 1.50% 1.50% 1.50% 1.50% 1.50% 1.50% EGTA 0.50% 1.00% Sodium decanoate 1.00% Sum 100.00% 100.00% 100.00% 100.00% 100.00% 100.00%

Anhydrous preparations 26 27 30 menthol 0.20% 0.20% 0.50% ethanol 2.00% 2.00% 1.50% Benzyl alcohol 8.00% 8.00% 1.00% Cetyl alcohol 2.00% Almond oil 5.00% 5.00% 3.00% lecithin 15.90% 15.90% 6.00% Lipmax 15.90% 15.90% 6.00% Propylene glycol 3.00% 3.00% 0.00% F127 Pluronic Powder 16.00% 16.00% 4.20% water 39.80% Sodium bicarbonate 26.00% 0.00% 28.00% Sodium carbonate 7.00% 33.00% 7.00% Durasoft 1.00% 1.00% 1.00% 100.00% 100.00% 100.00%

Example 2

Topical administration of bicarbonate

24 NCR nude 5 week old male mice were used in this study and were divided into 4 groups of 6 mice each. The topical composition was applied to the back of each mouse from hip to shoulder 3 times a day for 8 consecutive days for a total of 24 applications. The control group was administered orally with sodium bicarbonate dissolved in water. The groups are:

Transdermal formulations include penetrants to produce the following formulations: LIP-30.0%, EtOH-1.5%, BA-1.0%, menthol-0.5%, sodium bicarbonate-33.5%, PLU-F127-10.1%, PLU -Water-23.5%; That is, the formulation IB of Table 1. The concentration of sodium bicarbonate in the control group was 200 mM and the consumption was arbitrary. The concentration of sodium bicarbonate in the transdermal formulation was 33.5% by weight. Urine samples were collected 1, 3 and 6 hours after the first drug application and stored at 4 ° C for subsequent pH measurement. Urine was collected twice daily before the first application and 15 minutes after the last application on days 2-12. On day 8, mice were sacrificed 1 hour after the last drug application, and the back skin was harvested and placed on bibulous paper.

To set a baseline for pH without dosing, urine pH was measured at three time points during the day in seven mice at 09, 5 mice at 13 and 4 mice at 16:30, before starting the protocol. Did. Urine pH had an overall average value of 5.57 that did not change during this period.

As shown in Figure 1, all groups showed an increase in urine pH during the first 6 hours of treatment. The most significant increase occurred in group 3, administered with 220 microliters of the formulation.

This study was designed to be performed for 2 weeks (Monday to Friday), but was terminated after 8 days because members of Group 2 (1,110 microliters) developed skin irritation. This was also confirmed in the low-dose group receiving 30 microliters, namely Group 2.

Figure 2 shows the pH value of urine during the 8-day study process. Although there were some changes, the group receiving the highest dose (Group 4) was able to maintain a high pH during the course of the study.

The study was repeated using formulations 25, 28 and 29 of Table 6 with the results shown in Table 8 and formulations of Table 7 with the results shown in FIG. 3.

Average urine pH and total at the time of 2 collections on day 1 Urine 1 Piss 2 all SB water 6.30 6.03 6.15 25 6.36 6.77 6.62 28 6.92 6.86 6.89 29 6.82 7.31 6.97

As shown, transdermal administration was more effective than oral administration. Example 3

Percutaneous absorption in humans

Healthy human subjects aged 18-60 years were enrolled in a double-blind, placebo-controlled, randomized and cross-over in a designed study. Subjects applied the body weight of the formulations in Table 9 of 0.6 g / kg body weight per randomized group as follows: from leg to ankle to top of thigh; For the arm, from wrist to shoulder (including the triceps). Alternatively, 0.13 ounces per kg of body weight (approximately 8 ounces for a 140-pound subject) was ingested at 15 minutes, 1 hour 15 minutes, 2 hours 15 minutes, and 3 hours 15 minutes to control urine dilution.

LIP 30.0% ethanol 1.5% BA 1.0% Menthol (or limonene for lipophilicity) 0.5% Sodium bicarbonate 33.5% PLU-F127 10.1% PLU-water 23.5%

At time intervals (1, 2, 3 and 4 hours) from the start of application, subjects collected 10 to 20 ml (approximately one tablespoon) of urine and measured the pH. Table 10 shows the results.

Average urine pH value for each treatment and time point product Baseline
(n = 20) 1 hours
(n = 20) 2 hours (n = 20) 3 hours (n = 19) 4 hours (n = 20) Control (oral) 5.75 5.95 6.01 6.08 6.01 Formulation 5.86 6.11 6.27 6.24 6.23

No side effects were observed and transdermal preparations exceeded oral administration.

Example 4

Treatment of gout in humans

The topical application of sodium bicarbonate and menthol in transdermal delivery systems is intended to reduce pain associated with acute gout exposure to determine whether pain associated with acute gout flare can be effectively and safely reduced and the time to resolve can be reduced. A randomized, double-blind, placebo-controlled study of efficacy and safety of topical alkalizing treatment was performed.

Forty subjects over the age of 18 who had a history of gout and had gout that started within 36 hours in the hospital and were prescribed cochicine were randomly used in 20 groups.

The formulations used are shown in Table 11 below.

Gout preparation ingredient Contrast group Active group menthol 0.00% 0.50% ethanol 1.50% 1.50% Benzyl alcohol 1.00% 1.00% Cetyl alcohol 2.00% 2.00% Almond oil 3.00% 3.00% lecithin 10.00% 7.00% IPP 10.00% 7.00% Propylene glycol 5.00% 5.00% Poloxamer / Pluronic powder 9.00% 5.40% water 57.50% 33.60% Sodium bicarbonate 0.00% 33.00% Durosoft PK-SG 1.00% 1.00% 100.00% 100.00%

Subjects applied 10 ml of control or active cream to the entire limb of each affected joint three times a day. One identified "target joint" followed by up to two additional joints. Subjects reported joint pain measured using an 11-point scale (0 to 10, 0 to "no pain", 10 to "worst possible pain"). For the active group (two subjects), the pain reduction in the target joint was -3.0 points after 30 minutes and 24 hours, -3.5 points on days 4 and 6. Subjects in the control group showed no reduction at 30 minutes and 2 days, and only a decrease of -1 after 24 hours and on days 4 and 6. For the active group, the maximum reduction in secondary joint pain was -2 points, with little or no pain reduction compared to the control group.

Pain reduction associated with active product use was observed 15 minutes early after product application.

Example 5

Treatment of dermatosis in humans

1) tranexamic acid (TA), 2) TA and 5% glycolic acid ((GA), and 3) evaluating the efficacy and safety of TA used in combination with retinoic acid (RA) to modulate the improvement of dermaemia To do this, we conducted a randomized double-blind study evaluating tranexamic acid alone or in combination with a skin turnover agent in the transdermal delivery system to improve dermoderma. All formulations, including the control formulation, are alkaline. The formulations used are shown in Table 12.

Black dermatitis preparations ingredient contrast Active (TA) Active (TA + GA) Tranexamsan 0.000% 6.000% 6.000% Glycolic acid 0.000% 0.000% 5.000% Sodium carbonate 1.060% 1.000% 1.000% Triethanolamine 0.000% 0.000% 1.000% lecithin 13.300% 12.500% 12.500% IPP 13.300% 12.500% 12.500% Benzyl alcohol 1.060% 1.000% 1.000% Durosoft PK-SG 1.060% 1.000% 1.000% Poloxamer / Pluronic powder 9.255% 8.700% 6.900% water 60.745% 57.100% 52.900% Crisp Fruit Fragrance 0.220% 0.200% 0.200% 100.000% 100.000% 100.000%

Thirty-six women aged between 18 and 65, with moderate to severe dermatitis, were divided into 12 groups, respectively, and the subject was applied the formulation to the dermatosis site twice a day. The groups are as follows: Group 1: Control formulation; Group 2: Tranexamic acid (TA) 6% formulation; Group 3: TA 6% + glycolic acid 5% (GA) combination formulation; Group 4: TA 6% formulation and retinoic acid 0.25% (RA) (separate product, TA applied just prior to RA). Result measurements are standardized 2D with 5 views front, 45 degrees and 90 degrees. It was done by photography and included an Investigator global assessment of improvement compared to baseline, subject self-assessment of improvement compared to baseline and VISIA system photography (one site). Side effects were also evaluated.

Follow-up was done by calling on Day 3 and visiting at 2, 4, 8 and 12 weeks. In all of these groups, including the control group, 50% to 65% of subjects showed a reduction in pigmentation after 30 days by both independent and self-assessment. The results indicated that Group 2 generally improved for Groups 3 and 4, with 50% improvement in subjects in Group 2 and 30% and 65% in Groups 3 and 4, respectively, in a 30-day independent evaluation. Has been improved. (Self-evaluation results showed little difference at about 60%.)

Claims (24)

A method for inhibiting metastasis of a solid tumor in a subject, comprising administering parenterally non-systemic administration of an effective amount of a metastasis inhibitor to a subject in need thereof.
The method of claim 1, wherein the metastasis inhibitor is a buffer sufficient to increase the pH of the microenvironment of the tumor, a protease inhibitor, an inhibitor of Na ⁺ / H ⁺ exchange activity, an inhibitor of epidermal growth factor receptor (EGFR), a homologous region of src. A method selected from the group consisting of inhibitors of 2-containing protein tyrosinase phosphatase (Shp2), Wittaferrin A or combinations thereof.
The method of claim 2, wherein the administration is performed by contacting the subject's intact skin with an agent containing the metastasis inhibitor to provide the metastasis inhibitor to the subject through substantially intact or polished skin.
4. The method of claim 3, wherein the administration is to an intact skin and the formulation contains a penetrant that affects the skin penetration properties of the metastasis inhibitor.
5. The method of claim 4, wherein the penetrant comprises an amount of alcohol to provide 0.5 to 20% by weight of the weight of the formulation, and an amount of zwitterion to provide 25 to 70% by weight of the weight of the formulation. Way.
The penetrating agent of claim 4, wherein the penetrant comprises an amount of alcohol to provide 0.5 to 20% by weight of the weight of the formulation, and an amphoteric ion to provide 1 to 25% by weight of the weight of the formulation. Way.
7. The method of claim 6, wherein the penetrant further comprises a nonionic surfactant in an amount to provide 1 to 25% by weight of the weight of the formulation and a polar solvent in an amount of at least a molar excess of the nonionic surfactant or anhydrous (anhydrous).
The method of claim 7, wherein the nonionic surfactant is a poloxamer and the polar solvent is water.
The method of claim 4, wherein the penetrant is a formulation obtained,
i) an approximately 1: 1: 1 equimolar mixture of bile salt: lecithin: amphiphilic compound;
ii) at least one electrolyte sufficient to impart viscous and viscoelasticity to the formulation;
iii) one or more surfactants; And
iv) benzyl alcohol or analogs thereof;
How to include.
5. The method of claim 4, wherein the penetrant reduces thiol bonds, breaks hydrogen bonds and / or affects keratin dissolution; And / or
The penetrant further comprises at least one skin permeant (SPP); And / or
The penetrant further comprises a penetration enhancer; And / or
The preparation method comprising colloidal particles.
4. The method of claim 3, wherein the contact is contact with polished skin, and the formulation is applied under a sealed patch.
12. The method of claim 11, wherein the skin is polished by a microneedle device or sandpaper equivalent.
The method according to any one of claims 1 to 12, wherein the metastasis inhibitor comprises a buffer.
The method of claim 13, wherein the buffer is a carbonate buffer, a phosphate buffer, a Tris buffer or a chloroacetate buffer.
The method of claim 13, further comprising examining the subject for sensitivity or resistance to pH adjustment.
16. The method of claim 15, wherein the test comprises evaluating biopsy cells of the tumor in culture.
The method according to any one of claims 1 to 12, wherein the metastasis inhibitor is
Wittapain A, and / or
Protease inhibitors, and / or
An inhibitor of Na ⁺ / H ⁺ exchange, and / or
Inhibitors of epidermal growth factor receptor (EGFR), and / or
A method comprising an inhibitor of the src homology region 2-containing protein tyrosinasephosphatase (Shp2).
The method of any one of claims 1 to 12, wherein the metastasis inhibitor comprises a combination of agents.
The method of any one of claims 1 to 12, wherein the subject is a human, veterinary subject, or animal model.
A method for treating gout or hyperpigmentation in a subject, comprising administering parenterally non-systemic administration of a sufficient amount of a buffer to raise the pH at the location of the gout or hyperpigmentation to a subject in need of such treatment. Way.
The method of claim 20, wherein the administration is performed by contacting the subject's intact skin with an agent containing the metastasis inhibitor to provide the buffer to the subject through substantially intact or polished skin.
22. The method of claim 21, wherein the administration is to intact skin and the formulation further comprises a penetrant that affects the skin penetration properties of the buffer.
21. The method of claim 20, wherein the contact is contact with polished skin and the formulation is applied under a hermetic patch.
24. The method of any one of claims 20-23, wherein the subject is a human, veterinary subject, or animal model.

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