Improved synthesis of boronated amino acid compositions comprising TC220 and TC221 for boron neutron capture therapy and methods thereof

Abstract

Disclosed herein are boronated amino acid compositions comprising tyrosine derivatives TC220 and TC221 and improved methods of making TC220 and TC221. Thus, the TC220 and / or TC221 can be expanded to commercial scale and administered to patients as neutron trapping agents, and methods of treating cancer, immune disorders, and other diseases by utilizing a neutron capture therapy pattern are provided.

Description

Cross-references to related applications

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 628,747, filed August 17, 2023, the contents of which are incorporated herein by reference in their entirety.

[0002] Claims of rights for inventions made under federal government-funded research not applicable. Technical Field

[0003] This invention relates to the field of boron neutron capture therapy (BNCT). Specifically, it relates to improved methods for synthesizing boronized amino acids (a “BAA”) or (multiple “BAA”) compositions, including but not limited to BAAs denoted as TC220 and TC221, which can be used as mediators in human neutron capture therapy. The invention further relates to the treatment of cancer and other immune disorders and diseases. Background Technology

[0004] Cancer is the second leading cause of death worldwide, after coronary heart disease. Millions die from cancer each year, and in the United States alone, it kills more than half a million people annually, with 1,688,780 new cancer cases diagnosed in 2017 (American Cancer Society). While deaths from heart disease have been declining significantly, deaths from cancer are generally on the rise. Unless medical advancements reverse current trends, cancer is projected to become the leading cause of death by the beginning of the next century.

[0005] Several cancers have high mortality rates. Specifically, lung cancer (18.4% of all cancer deaths), breast cancer (6.6% of all cancer deaths), colorectal cancer (9.2% of all cancer deaths), liver cancer (8.2% of all cancer deaths), and stomach cancer (8.2% of all cancer deaths) are the leading causes of cancer death in both sexes across all age groups worldwide (GLOBOCAN 2018). These cancers share a common lethal characteristic with almost all other cancers: metastasis to sites far from the primary tumor, and with very few exceptions, metastatic disease is fatal. Furthermore, even for those cancer patients who initially survive their primary cancer, the shared experience has shown that their lives are significantly altered. Many cancer patients experience intense anxiety due to the awareness of potential recurrence or treatment failure. Many cancer patients also experience physical weakness after treatment. In addition, many cancer patients experience disease recurrence.

[0006] Despite advancements in cancer treatments and improved survival rates over the past few decades, the heterogeneity of cancer necessitates novel treatment strategies utilizing multiple modalities. This is particularly true for solid tumors located in anatomically critical sites (e.g., glioblastoma, head and neck squamous cell carcinoma, and lung adenocarcinoma) where standard radiotherapy and / or chemotherapy are sometimes limited. However, the detrimental effects of these therapies include chemoresistance and radiation resistance, which, in addition to serious side effects that reduce patients' quality of life, promote localized recurrence, distant metastasis, and secondary primary tumors.

[0007] Neutron capture therapy (NCT) is a promising form of radiation therapy. It is a technique that uses boron compounds to selectively kill tumor cells while preserving normal cells. BNCT relies on non-radioactive... 10 The tendency of B isotopes to absorb superthermal neutrons, which fall within 0.5 keV. <E n In the low-energy range of <30 keV, after neutron capture, boron atoms undergo nuclear fission, producing alpha particles and recoiled lithium nuclei. 7 Li), as shown below:

[0008] 10 B + n→ 7 Li + 4 He

[0009] Alpha particles deposit high-energy particles (150 keV / μm) along a short path that is essentially confined to the diameter of a single cell, causing double-strand DNA breaks, which subsequently lead to cancer cell death through apoptosis. Therefore, BNCT integrates the concepts of chemotherapy and targeted therapy with the gross anatomical localization of traditional radiotherapy.

[0010] Although the conceptual technologies of NCT and specifically boron neutron capture therapy (BNCT) are well-known, technological limitations associated with this type of treatment have slowed progress. In early studies conducted in the 1960s using MIT's research reactor, dozens of patients were treated with disodium decaborate, which was thought to be less toxic than previously used simple boron compounds but capable of delivering more boron to cells. Unfortunately, BNCT research in the United States was halted due to severe brain necrosis in patients undergoing BNCT and the potential hazards of using nuclear reactors.

[0011] In 1968, Hiroshi Hatanaka re-examined the clinical application of boron carbamate (BSH) in BNCT in Japan by directing the beam to surgically exposed intracranial tumors, reporting a 5-year survival rate of 58%. In 1987, Japanese clinicians used boron phenylalanine (BPA) as a boron compound in BNCT to treat malignant melanoma. Thus, BNCT experienced a slow resurgence, albeit limited to countries with access to research reactor facilities capable of delivering superthermal neutron beams. Currently, NCT treatment is experiencing a resurgence due to technological improvements in (i) the infusion and delivery of trapping compounds preferably focused on the tumor, and (ii) the availability and abundance of neutron beams through cyclotrons.

[0012] Proton-boron fusion reactions depend on naturally abundant 11 B isotopes, rather than those required for BNCT. 10 B isotopes. Unlike BNCTs, protons ( 1 H) and boron ( 11 B) The fusion reaction between nuclei will emit three alpha particles: p+ 11 B → 3α. Proton beams have the advantage of Bragg-peak characteristics, which can reduce damage to normal tissues, and when combined with proton capture, they can enhance the efficacy of proton therapy on their own.

[0013] Boron loaders have been evolving since the 1950s and were reviewed in Nedunchezian et al., Journal of Clinical and Diagnostic Research, Vol. 10(12) (December 2016). In short, first-generation boron compounds, represented by boric acid and its derivatives, were either toxic or had low tumor accumulation / retention. BPA and BSH are considered second-generation compounds that emerged in the 1960s. They exhibit significantly lower toxicity and better PK and biodistribution. The BPA-fructose complex is considered a third-generation compound and has been used since 1994 in patients with H&N, glioblastoma, and melanoma treated with BNCT. BPA-fructose and BSH are by far the only compounds used clinically as boron loaders, although tumor targeting of low and high molecular weight biomolecules (such as nucleosides, porphyrins, liposomes, nanoparticles, and mAbs) has been evaluated in preclinical models. The main drawback of BPA-fructose is its relatively low solubility, coupled with its rapid clearance, which prevents it from reaching high or peak serum doses (C60) in the blood. max This is one of the driving factors affecting tumor uptake.

[0014] Based on the foregoing, it will be apparent to those skilled in the art that new therapeutic paradigms are needed in the treatment of cancer and immune diseases. New disease treatments can be achieved through the use of modern chemical synthesis and boron-modified natural amino acids, with the overall goal of more effective treatment, fewer side effects, and lower production costs.

[0015] In view of the current deficiencies associated with NCT, the object of the present invention is to provide a new and improved method for treating one or more cancers, immune disorders and other diseases using boronized amino acids and NCT. Summary of the Invention

[0016] This invention provides compositions comprising natural amino acids that have been boronized through chemical synthesis for use as a delivery modulus for treating human diseases such as cancer, immune disorders (including but not limited to rheumatoid arthritis and ankylosing spondylitis), and other cellular diseases (including but not limited to Alzheimer's disease). In some embodiments, the boronized amino acids comprise naturally occurring amino acids such as phenylalanine, tryptophan, tyrosine, histidine, and any other naturally occurring amino acids shown in Table I.

[0017] In another embodiment, the present invention includes TC220.

[0018] In another embodiment, the present invention includes TC221.

[0019] In another embodiment, the present invention includes an improved method for synthesizing TC220.

[0020] In another embodiment, the present invention includes an improved method for synthesizing TC221.

[0021] In another embodiment, the present invention includes a method for concentrating boron in cells, the method comprising (i) synthesizing boronized amino acids (“BAA”); (ii) administering the BAA to a patient; and (iii) irradiating the cells with neutrons.

[0022] In another embodiment, the present invention includes a method for concentrating boron in cells, the method comprising (i) synthesizing TC220, (ii) administering the TC220 to a patient, and (iii) irradiating the cells with neutrons.

[0023] In another embodiment, the present invention includes a method for concentrating boron in cells, the method comprising (i) synthesizing TC221, (ii) administering the TC221 to a patient, and (iii) irradiating the cells with neutrons.

[0024] In another embodiment, this disclosure teaches a method for synthesizing BAA.

[0025] In another embodiment, this disclosure teaches methods for treating cancer, immune disorders, and other diseases in humans. Attached Figure Description

[0026] Figure 1 Chemical structure of TC220 hydrochloride.

[0027] Figure 2 Chemical structure of TC221.

[0028] Figure 3 Synthesis scheme of TC220 hydrochloride.

[0029] Figure 4 Preparation of (S)-2-(tert-butoxycarbonylamino)-3-(4-hydroxy-3-iodophenyl)propionic acid.

[0030] Figure 5 Preparation of trimethylsilyl ester (S)-2-(tert-butoxycarbonylamino)-3-[3-iodo-4-(trimethylsiloxy)phenyl]propionate.

[0031] Figure 6 . 10 Preparation of B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionic acid.

[0032] Figure 7 . 10 Preparation of B-TC220 hydrochloride.

[0033] Figure 8 . 10 Purity curve of B-TC220 hydrochloride. Figure 8 (A) shows the wavelengths and mass spectra (TICs) for all analyses. Figure 8 (B). This shows the positive M / Z of M+1 = 346.23. Figure 8 (C) shows the UV spectrum at 225 nm. Figure 8 (D). Shows the percentage of the area of ​​the detectable UV peaks.

[0034] Figure 9 Synthetic scheme for converting TC220 hydrochloride to TC221.

[0035] Figure 10 Used to 10 B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionic acid is converted to 10 Synthetic scheme for methyl B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionate.

[0036] Figure 11 Chemical synthesis of N-Boc-Tyr(3-B(OH)2, 4-OMe)-OMe to Boc-Tyr(3-B(OH)2, 4-OMe)-OH.

[0037] Figure 12 Chemical synthesis of Boc-Tyr(3-B(OH)2, 4-OMe)-OH to TC221. Detailed Implementation

[0038] Chapter Summary

[0039] I.) Definition

[0040] II.) BPA

[0041] III.) BSH

[0042] IV.) Boron

[0043] a. Boron, generally

[0044] V.) Naturally occurring amino acids

[0045] VI.) Borated amino acids (BAA)

[0046] a. Amino acid composition

[0047] b. BAAs containing tyrosine (TC220 and TC221)

[0048] c. New and improved synthesis of TC220 and TC221

[0049] VII.) Boron neutron capture therapy using TC220 and TC221

[0050] VIII.) Proton-boron fusion therapy using TC220 and TC221

[0051] IX.) Methods for delivering TC220 and TC221 into cells

[0052] X.) Reagent kits / products

[0053] I.) definition:

[0054] Unless otherwise defined, all technical terms, symbols, and other scientific terms or expressions used herein are intended to have the meaning commonly understood by one of ordinary skill in the art to which this invention pertains, unless the context clearly indicates otherwise. In some instances, terms having a commonly understood meaning are defined herein for clarity and / or ease of reference, and the inclusion of these definitions herein should not necessarily be construed as indicating a substantial difference from the meaning commonly understood in the art.

[0055] When a trade name is used in this document, unless the context otherwise indicates, reference to a trade name also refers to the product formulation of the product in which the trade name is used, generic drugs, and one or more active pharmaceutical ingredients.

[0056] The terms “advanced cancer,” “locally advanced cancer,” “advanced disease,” and “locally advanced disease” refer to cancer that has extended beyond the capsule of the relevant tissue and are intended to include stage C disease according to the American Urological Association (AUA) system, stage C1-C2 disease according to the Whitmore-Jewett system, and stage T3-T4 and N+ disease according to the TNM (tumor, nodule, metastasis) system. Generally, surgery is not recommended for patients with locally advanced disease, and outcomes are significantly worse compared to patients with clinically localized (organ-specific) cancer.

[0057] "Amino acid" refers to a simple organic compound containing both a carboxyl group (-COOH) and an amino group (-NH2).

[0058] "Boronization" refers to the reaction that produces organoboron compounds through the functionalization of aliphatic and aromatic CH bonds.

[0059] "Boronized amino acid" (BAA) refers to a compound containing naturally occurring amino acids, such as those shown in Table I, which have undergone a boronization reaction. BAAs can be synthesized in various forms, depending on the base amino acid used.

[0060] "TC220" means containing Figure 1 The compound with the chemical structure shown. For the purposes of this disclosure, TC220 may be referred to as "TC220" or "TC220 hydrochloride".

[0061] "TC221" means containing Figure 2 The compound with the chemical structure shown.

[0062] The term "compound" means and covers chemical compounds (e.g., BAA) themselves, and excludes, whether explicitly stated otherwise and unless the context clearly states otherwise: amorphous and crystalline forms of the compound, including polycrystalline forms, wherein these forms may be part of a mixture or exist alone; free acid and free base forms of the compound, which are generally those shown in the structures provided herein; isomers of the compound, which mean optical isomers and tautomers, wherein optical isomers include enantiomers and diastereomers, chiral isomers and achiral isomers, and optical isomers include isolated optical isomers and mixtures of optical isomers, including racemic and non-racemic mixtures, wherein the isomers may be in isolated forms or mixtures with one or more other isomers; isotopes of the compound, including deuterium-containing isomers. Compounds of tritium, including compounds containing radioactive isotopes, including therapeutically and diagnostically effective radioactive isotopes; polymeric forms of compounds, including dimers, trimers, etc.; salts of compounds, preferably pharmaceutically acceptable salts, including acid addition salts and base addition salts, including salts having organic and inorganic counterions, and including zwitterionic forms, wherein if a compound associates with two or more counterions, the two or more counterions may be the same or different; and solvates of compounds, including hemisolvents, monosolvents, and disolvents, including organic and inorganic solvates, the inorganic solvates including hydrates, wherein if a compound associates with two or more solvent molecules, the two or more solvent molecules may be the same or different. In some cases, references to compounds of the present invention herein will include explicit references to one or more of the above forms, such as salts and / or solvates; however, such references are for emphasis only and should not be construed as excluding other forms identified above.

[0063] As used herein, the term “inhibition” or “inhibition of…” means to reduce a measurable amount or to prevent it entirely.

[0064] The term "mammal" refers to any organism classified as a mammal, including mice, rats, rabbits, dogs, cats, cows, horses, and humans. In one embodiment of the invention, the mammal is a mouse. In another embodiment of the invention, the mammal is a human.

[0065] The terms “metastatic cancer” and “metastatic disease” refer to cancer that has spread to regional lymph nodes or distant sites and include stage D disease according to the AUA system and stage T×N×M+ disease according to the TNM system.

[0066] "Molecular recognition" refers to a chemical event in which a host molecule can form a complex with a second molecule (i.e., the guest). This process occurs through non-covalent chemical bonds, including but not limited to hydrogen bonding, hydrophobic interactions, and ionic interactions.

[0067] "Pharmaceutical acceptable" means a non-toxic, inert, and / or composition that is physiologically compatible with humans or other mammals.

[0068] The term "neutron trapper" refers to a stable, non-reactive chemical isotope that produces alpha particles when activated by a neutron.

[0069] The term "neutron capture therapy" refers to a non-invasive treatment modality used to treat locally invasive malignancies such as primary brain tumors and recurrent head and neck cancers, as well as other immune disorders and diseases, by irradiating a neutron capture agent with neutrons.

[0070] As used herein, the terms “for treatment” or “therapeutic” and grammatically related terms refer to any improvement in any outcome of the disease, such as increased survival, reduced morbidity, and / or lessened side effects, which are byproducts of alternative treatment modalities; as is readily understood in the art, complete eradication of the disease is preferred, but nevertheless is not a requirement of therapeutic action.

[0071] II.) BPA

[0072] For reference and in the context of existing technology, ( 10 B)-BPA, L-BPA, or 4-dihydroxyboryl-L-phenylalanine (Sigma Aldrich, St. Louis, MO) is a compound with the chemical formula C9H 12 Synthetic compounds of BNO4. The structure is shown below:

[0073]

[0074] It is also an important boron compound that can be used to treat cancer via BNCT. It is a widely known compound, and many synthetic methods have been developed (see, US 8,765,997, Taiwan Biotech Co, Ltd., Taoyuan Hsein, Taiwan, China, and US2017 / 0015684, Stella Pharma Corp., Osaka Prefecture Univ., Osaka, Japan).

[0075] III.) BSH

[0076] In addition to BPA, BSH or boraxane or BSH boraxane or boraxane 10 B, or undecylhydro-closed-dodecylborylthiol, is a known chemical formula of Na₂B. 12 H 11 Synthetic compounds of SH. The structure is shown below:

[0077]

[0078] Boron atoms are represented by points at the vertices of an icosahedron. BSH is used as a trapping agent in BNCT. Generally, BSH is injected intravenously and becomes concentrated in tumor cells. The patient then receives radiation therapy with atomic particles called neutrons. The neutrons fuse with the boron nuclei in the BSH and produce high-energy alpha particles that kill the tumor cells.

[0079] IV.) Boron

[0080] (a.) Boron, generally

[0081] Generally, and for the purposes of this disclosure, boron is a chemical element having the symbol B and atomic number 5. Natural boron is primarily used in compounds consisting of two stable isotopes, boron-10 and boron-11. The boron-10 isotope can be used to capture superthermal neutrons, making it a promising tool in the context of treatments using boron neutron capture therapy. Biologically, the boron compounds disclosed herein are non-toxic to humans and animals. Based on the foregoing, it will be apparent to those skilled in the art that improved modalities for delivering high concentrations of boron to cancer cells are advantageous. The purpose of this disclosure is to provide these advantages.

[0082] V.) Naturally occurring amino acids

[0083] Generally, and for the purposes of this disclosure, naturally occurring amino acids are organic compounds containing amine (-NH2) and carboxyl (-COOH) functional groups, as well as a side chain (R group) characteristic of each amino acid. The key elements of amino acids are carbon (C), hydrogen (H), oxygen (O), and nitrogen (N), although other elements have been found in the side chains of some amino acids. Approximately 500 naturally occurring amino acids are known (although only 20 appear in the genetic code (Table I)) and can be classified in many ways. They can be classified according to the position of their core structural functional groups as α- (alpha-), β- (beta-), γ- (gamma-), or δ- (delta-) amino acids; other categories involve polarity, pH level, and the type of side chain group (aliphatic, acyclic, aromatic, hydroxyl-containing, or sulfur-containing, etc.). In the form of proteins, amino acid residues form the second largest component (water is the largest) of human muscle and other tissues. In addition to their role as residues in proteins, amino acids participate in many processes, such as neurotransmitter transport and biosynthesis.

[0084] The twenty (20) amino acids directly encoded by the genetic code (see Table I) can be grouped into several groups based on their properties. The main factors are charge, hydrophilicity or hydrophobicity, size, and functional groups. These properties are important for protein structure and protein-protein interactions. Water-soluble proteins tend to bury their hydrophobic residues (Leu, Ile, Val, Phe, and Trp) in the middle of the protein, while their hydrophilic side chains are exposed to aqueous solvents.

[0085] Integrated membrane proteins often have an outer ring of exposed hydrophobic amino acids that anchor them to the lipid bilayer. Between these two extremes, some peripheral membrane proteins have a plate of hydrophobic amino acids on their surface that are locked to the membrane. Similarly, proteins that must bind to positively charged molecules have surfaces rich in negatively charged amino acids, such as glutamic acid and aspartic acid, while proteins that bind to negatively charged molecules have surfaces rich in positively charged chains, such as lysine and arginine. These differ in the hydrophobicity levels of the amino acid residues.

[0086] Some amino acids have special properties, such as cysteine, which can form covalent disulfide bonds with other cysteine ​​residues; proline, which forms a cycle with the polypeptide backbone; and glycine, which is more flexible than other amino acids.

[0087] VI.) Borated amino acids (BAA)

[0088] By briefly introducing and better understanding the background of the inventive efforts of this disclosure, it is noted that large neutral amino acid transporter 1 (LAT-1, SLC7a5) is a sodium- and pH-independent transporter that delivers essential amino acids (e.g., leucine, phenylalanine) to cells. The functional transporter is a heterodimeric disulfide-bonded complex composed of the multi-transmembrane subunit SLC7a5 and the single-transmembrane subunit SLC3a2 (CD98). LAT-1 is a major transporter that guides essential amino acids across compartments such as the placenta or blood-brain barrier. In addition, LAT-1 also transports thyroid hormones T3 and T4 (see FRIESEMA et al., Endocrinology, 142(10): 4339-4348 (2001)), the dopamine precursor L-DOPA, and amino acid-related exogenous compounds such as the drugs melphalan and gabapentin (see Uchino et al., Molecular Pharmacology, 61:729-737 (2002)). Furthermore, its expression is highly upregulated in several types of human cancers, characterized by a strong metabolic and growth demand for amino acids (see SINGH et al., International Journal of Molecular Sciences, 2018, 19, 1278). Furthermore, it has been reported that the properties of the amino acid side chains affect the selectivity of LAT-1 for various amino acids, with the following order in terms of increasing transport rate: Phe > Trp > Leu > Ile > Met > His > Tyr > Val (see Kanai et al., Journal of Biochemistry, Vol. 273, No. 37, pp. 23629–23632 (1998)). However, the effects of additional boron modifications on amino acids are unknown in the art, and this disclosure represents a groundbreaking breakthrough.

[0089] The therapeutic potential of BNCT as an effective cancer treatment lies in the sufficient amount of [unclear text - possibly a typo, should be 6] within cancer cells. 10 Selective accumulation of B.

[0090] Based on the foregoing, those skilled in the art have demonstrated that essential amino acid transporters (such as LAT1) are responsible for the uptake of certain naturally occurring amino acids. See Scalise et al., Frontiers in Chemistry, Vol. 6, No. 243 (June 2018). In light of this principle, this disclosure envisions synthesizing naturally occurring amino acids via borylation reactions to produce borated amino acids (“BAA”) with tumor-finding and tumor-localizing properties, for use as neutron trapping agents in boron neutron capture therapy (“BNCT”) and / or boron proton capture therapy (commonly referred to as proton-boron fusion therapy (“PBFT”)). See, for example, Hattori et al., Journal of Medicinal Chemistry, 55, 6980-6984 (2012).

[0091] (a) amino acid composition

[0092] In another embodiment, BAA having the following formula is within the scope of this disclosure (“tyrosine derivative”):

[0093]

[0094] in:

[0095] E = CO2H, CONHB 12 H 11 、B(OH)2; and

[0096] X = H, B(OH)2, Bpin, (-O-CH2CH2)2-OB 12 H 11 Or BF3 - .

[0097] In another embodiment, tyrosine derivatives having the following formula are within the scope of this disclosure:

[0098]

[0099] in:

[0100] R = H, CH3, or CF3;

[0101] X1 = H, B(OH)2 or BF3 - ;and

[0102] X2 = H, B(OH)2 or BF3 - .

[0103] (b) BAA containing tyrosine (TC220 hydrochloride and TC221)

[0104] Tyrosine is an essential amino acid with the following chemical formula:

[0105]

[0106] Tyrosine is known to readily cross the blood-brain barrier. Once in the brain, it is a precursor to the neurotransmitters dopamine, norepinephrine, and epinephrine (more commonly known as adrenaline). These neurotransmitters are an important part of the body's sympathetic nervous system, and their concentrations in the body and brain are directly dependent on dietary tyrosine. Tyrosine metabolism is rapid. Folic acid, copper, and vitamin C are cofactor nutrients for these responses. Tyrosine is also a precursor to hormones, the thyroid gland, catechol estrogens, and the major human pigment, melanin. Tyrosine is an important amino acid in many proteins, peptides, and even enkephalins, the body's natural pain relievers. Valine and other branched-chain amino acids, and possibly tryptophan and phenylalanine, may reduce tyrosine absorption. Several genetic errors can occur in tyrosine metabolism, such as Hawkins' nuria and hypertyrosinemia I. The most common is increased tyrosine levels in the blood of premature infants, manifested as decreased motor activity, lethargy, and poor feeding. Infections and intellectual impairment may occur. Some adults may also experience elevated tyrosine levels in their blood. This indicates a need for more vitamin C. Generally, tyrosine is needed under stress, and tyrosine supplements can prevent stress-induced norepinephrine depletion and may even cure biochemical depression.

[0107] In addition, various derivatives of tyrosine have been evaluated as tracers for whole-body imaging using PET, and some of them have been approved for specific indications, including neuroendocrine disorders and cancer. These derivatives include... 18 F-fluorine-L-DOPA (DOI: 10.2967 / jnumed.114.145730), 18 F-Fluoro-L-α-methyltyrosine (FAMT) (Inoue, Journal of Nuclear Medicine 1998; 39:663-667 and 10.2967 / jnumed.112.103069).

[0108] In addition, Ishiwata et al. described O-[ 18 F]Fluoromethyl-L-tyrosine (e.g., 18F-FMT was investigated, and its biodistribution in rats bearing hepatocellular carcinoma was examined. See Nuclear Medicine and Biology 31 (2004) 191–198. As shown, the tracer accumulated in the pancreas and achieved meaningful contrast at sixty (60) minutes, thus providing visualization of the tumor. However, some degree of defluorination was observed, which may be due to tracer uptake in the bone marrow. 18 F-FMT defluorination and 18 F-fluoroethyltyrosine (i.e.) 18 A contrast-enhanced non-defluorinated formation of F-FET (an approved imaging tracer for advanced gliomas) (see 10.2967 / jnumed.114.140608 and NCT04001257). Notably, the increased uptake of the aforementioned PET tracer in gliomas and other tumors is mediated by LAT-1, the same large neutral amino acid transporter that mediates BPA uptake into head and neck, GBM, and melanoma lesions.

[0109] Based on the foregoing, this disclosure focuses on the synthesis of boronized tyrosine analogs to evaluate LAT-1 expression in selected cell lines and demonstrate that these boronized amino acid analogs exhibit uptake in cancer cell lines. Furthermore, this disclosure demonstrates that tyrosine analogs are absorbed in a dose-dependent manner by FaDu-based xenografts in immunodeficient mice.

[0110] Therefore, this disclosure envisions the use of borotyrosine as a neutron trap in certain cancers.

[0111] In one embodiment of this disclosure, the BAA containing tyrosine is represented as TC220 hydrochloride and has the following characteristics: Figure 1 The following chemical formulas are shown.

[0112] In one embodiment of this disclosure, the BAA containing tyrosine is denoted as TC221 and has the following characteristics: Figure 2 The following chemical formulas are shown.

[0113] The synthesis of TC220 and TC221 is challenging due to the hydroxyl group of tyrosine adjacent to boric acid. This hydroxyl group is an electron-donating group and hinders the pinacol-borane deprotection step in the synthesis. Consequently, known synthesis of TC220 and TC221 results in low yields and difficulty in removing impurities. Therefore, the object of this disclosure is to provide a novel synthesis of TC220 and TC221 that yields high yields and purity on a scale of up to one (1) gram.

[0114] As background, TC220 and TC221 are highly soluble in water. However, unlike BPA, the only currently approved boron carrier for BNCT, TC220 does not require fructose to aid dissolution. Additionally, TC221 does require fructose; however, its solubility threshold is much higher than that of BPA. Therefore, using TC220 or TC221 allows for the application of higher concentrations and smaller volumes of boron compounds compared to BPA, which is currently feasible with L-BPA fructose (or sorbitol) formulations. The clinical significance of achieving higher boron concentrations in tumors will translate into more potent neutron irradiation in BNCT and / or PBFT therapies, and ultimately, lower cancer recurrence rates.

[0115] Therefore, the purpose of this disclosure is to teach new and improved synthesis of TC220 and TC221.

[0116] (c) New and improved synthesis of TC220 and TC221

[0117] TC220 (e.g.) Figure 1 (as shown) and TC221 (as shown) Figure 2 The synthesis of (shown) is efficient on a laboratory scale. However, the formation of L-DOPA and Tyr has been observed to follow the conventional Miyaura coupling (i.e., Pd coupling of bis(pinacol)borane followed by NaIO4 deprotection). This results in a major byproduct, which requires chromatographic removal.

[0118] Additionally, when the N or C ends are deprotected, TC220 (e.g.) will not function properly unless an excessive amount of BBr3 is added. Figure 1 (as shown) and TC221 (as shown) Figure 2 The synthesis shown will not proceed. However, this introduces another problem: the synthesis leads to rapid deboronization. It should also be noted that initiating the synthesis above 0°C will also result in deboronization.

[0119] Therefore, the purpose of this disclosure is to achieve a new and improved synthesis scheme for TC220 and TC221, thereby eliminating impurities generated by Pd coupling and reducing the number of steps. (See also...) Figure 3 ).

[0120] The preparation method of the newly synthesized material makes chromatography unnecessary for final purification. Figure 8 The chromatographic conditions specified in section A are used to analyze the purified target material, and TC220 (such as...) will be generated. Figure 1 (as shown) and TC221 (as shown) Figure 2 As shown), and allows commercial scaling up to one (1) gram.

[0121] In a preferred embodiment, the present invention comprises the synthesis of TC220 and TC221, which involves the conversion of L-tyrosine into the target material. The conversion of L-tyrosine into N-Boc-Tyr(3-Br, 4-MeO)-OMe was first presented in Ghosh, S. et al., ARKIVOC, 2009 (vii), 72-78.

[0122] The conversion of N-Boc-Tyr(3-Br, 4-MeO)-OMe to arylboronic acid N-Boc-Tyr(3-B(OH)2-4-MeO)-OMe is achieved by palladiumization followed by metal exchange to the desired boric acid composition. 30 mL of methanol and 12 mL of dimethoxyethane were charged into a flame-dried argon-quenched flask. 2.8 g of potassium acetate was added to the solution, followed by 5 g of Boc-Tyr(3-Br, 4-OMe)-OMe, and then 1.3 g of tetrahydroxyborane. Finally, catalytic Pd and 3 mg of chloro[(tri-tert-butylphosphine)-2-(2-aminobiphenyl)]palladium(II) were added to the reaction mixture. The reaction was stirred overnight at 20 °C under an argon atmosphere. Upon completion of the reaction, 20 mL of water was slowly added, and the reaction was quenched for 30 min. The solids were removed by filtration. The organic solvent was then removed under reduced pressure. The aqueous layer was then washed three times with ethyl acetate. The organic layers were combined and concentrated under reduced pressure. The crude material was further purified by rapid chromatography (on silica, in hexane containing 25% ethyl acetate). After removal of the organic solvent, the target material was separated as a white solid in 75% yield.

[0123] Following the introduction of boric acid, synthetic differentiation was performed to form compounds TC220 and TC221. The selective saponification of N-Boc-Tyr(3-B(OH)2-4-MeO)-OMe to N-Boc-Tyr(3-B(OH)2-4-MeO)-OH was highlighted in Gao Xi et al., ARKIVOC, 2009 (vii), 72-78. After saponification, tert-butyl carbamate was removed to reveal the structure of TC221. A solution of 4 M hydrochloric acid in dioxane was added to a flask containing 1 g of N-Boc-Tyr(3-B(OH)2, 4-OMe)-OH. After one hour, no Boc-protected starting material was observed. Volatile solvents and acids were removed under reduced pressure, and the target material was purified by preparative LC.

[0124] VII.) Boron neutron capture therapy using TC220 and TC221

[0125] One aspect of this disclosure is the use of TC220 and TC221 as modalities for boron neutron capture therapy (BNCT) and / or boron proton capture therapy (“BPCT”). Simply put, BNCT is a binary treatment modality in which neither component alone is lethal or toxic to the tumor. These two components comprise (i) the infusion or delivery of a capture compound preferentially concentrated in the tumor and (ii) irradiation of the tumor site by neutrons or by protons. In BNCT, thermal neutrons and… 10 The cross-section of the boron-boron interaction is relatively large; therefore, the boron nucleus splits into... 4 He 2+ and 7 Li + The possibility is higher. Considering He 2+ and Li + Due to the high ionization capacity and short travel distance of boron, cells preferably enriched with boron are killed, while healthy cells suffer much less damage due to the lack of high boron concentrations. With this in mind, the advantage of BNCT is the destruction of tumor cells without highly invasive surgical procedures. However, as those skilled in the art will understand, success depends on… 10 B exhibits high concentration and selective localization within tumor cells.

[0126] In one embodiment, 10 B concentrates on TC220 and / or TC221. Then TC220 and / or TC221 are given to the patient, and TC220 and TC221 are localized to the tumor cells. [The remaining text appears to be incomplete and requires further context.] 10 B's TC220 and TC221 were concentrated in the tumor, and the tumor was irradiated with superheated neutrons. The tumor cells were destroyed.

[0127] VIII. Proton-boron fusion therapy using BAA

[0128] Another aspect of this disclosure is the use of TC220 and TC221 as a modality for proton-boron fusion therapy (PBFT). Simply put, the proton-boron fusion reaction was introduced in the 1960s. In proton ( 1 H) and boron particles ( 11Following the reaction between B), three types of alpha particles are emitted. These three alpha particles damage tumor cells, just like the alpha particles in BNCT. Theoretically, in the case of PBFT, the therapeutic effect of each incident particle is three times (3x) that of BNCT. Furthermore, because of the Bragg peak characteristics of the proton beam, damage to normal tissue can be reduced. In general, many studies have been conducted on the use of alpha particles to treat tumors. Two key points should be considered for dose delivery using alpha particles. First, boron uptake should be accurately labeled to target cells. As mentioned earlier, alpha particles are generated where borate compounds accumulate. If this occurs in normal tissue near the tumor area, then the alpha particles will damage both normal tissue and tumor cells. Second, the number of alpha particles generated is also an important factor for effective therapy. By using PBFT, a more effective therapy can be achieved compared to BNCT alone or conventional proton therapy.

[0129] In one embodiment, 10 B and / or 11 B concentrates on TC220 and TC221. Then, TC220 and TC221 are given to the patient, and TC220 and TC221 are localized to the tumor cells. [The remaining text appears to be incomplete and requires further context.] 10 B and / or 11 B's TC220 and TC221 were concentrated in the tumor, and the tumor was irradiated with superheated neutrons. The tumor cells were destroyed.

[0130] IX. Methods for delivering TC220 and TC221 into cells

[0131] As those skilled in the art will understand, the ability to efficiently deliver high concentrations of boron into cells is an advantage of the present invention.

[0132] The results show that the TC220 and TC221 of this disclosure enable the safe application of higher amounts of boron to mammalian cells. In short, the TC220 and TC221 of this disclosure are prepared as shown in this disclosure. The resulting TC220 and TC221 are taken up by tumor cells via upregulated LAT-1 transporter.

[0133] X.) Reagent kits / products

[0134] For use in the laboratory, prognostic, preventative, diagnostic, and therapeutic applications described herein, the kits are within the scope of this invention. Such kits may comprise carriers, packages, or containers separated to house one or more containers (e.g., vials, tubes, etc.), each container containing a single element for use in the methods described, and a label or insert containing instructions for use as described herein. For example, the container may contain one or more TC220s and TC221s of this disclosure. The kit may comprise containers including pharmaceutical units. The kit may include all or part of one or more TC220s and TC221s and / or diagnostic assays for detecting cancer and / or other immune disorders.

[0135] The kits of the present invention will typically include the aforementioned container and one or more other associated containers containing materials desirable from a commercial and user perspective, including: buffers, diluents, filters, needles, syringes; carriers, packaging, containers, vials and / or tube labels listing contents and / or instructions for use; and packaging inserts with instructions for use.

[0136] Labels may be presented on or with the container to indicate that the composition is intended for a specific therapeutic or non-therapeutic application, such as prognostic, preventative, diagnostic, or laboratory use, and may also indicate in vivo or in vitro usage guidelines, as described herein. Guidelines and / or other information may also be included in inserts or labels included with or on the kit. Labels may be on or associated with the container. A label may be on the container when the letters, numbers, or other characters forming the label are molded or etched into the container itself; a label may be associated with the container when it is present within a receiver or carrier that also contains the container, such as as a packaging insert. Labels may indicate that the composition is intended for the diagnosis, treatment, prevention, or prediction of conditions such as cancer or other immune disorders.

[0137] The terms "reagent kit" and "product" can be used synonymously.

[0138] In another embodiment of the invention, one or more articles contain compositions such as one or more TC220 and TC221 of this disclosure. The articles typically include at least one container and at least one label. Suitable containers include, for example, bottles, vials, syringes, and test tubes. Containers can be formed from various materials such as glass, metal, or plastic. Containers can contain one or more TC220 and TC221 and / or one or more therapeutic doses of TC220 and TC221.

[0139] The container may also alternatively contain compositions effective for the treatment, diagnosis, prediction, and prevention of symptoms and may have a sterile access port (e.g., the container may be an intravenous solution bag or vial with a stopper that can be punctured by a hypodermic needle). The active agents in the composition may be TC220 and TC221 of this disclosure.

[0140] The article may further comprise a second container containing a pharmaceutically acceptable buffer solution, such as phosphate-buffered saline, Ringer's solution, and / or dextrose solution. The article may further comprise other materials desirable from a commercial and user perspective, including other buffers, diluents, filters, stirrers, needles, syringes, and / or packaging inserts with usage instructions and / or usage guidelines.

[0141] Exemplary Examples

[0142] 1) A composition comprising the following chemical structure:

[0143]

[0144] 2) A composition comprising the following chemical structure:

[0145]

[0146] 3) A composition produced by a method of converting L-tyrosine into a target material, said method comprising the following synthesis:

[0147]

[0148] 4) The method of claim 3, wherein the resulting composition is... Figure 1 As shown in the image.

[0149] 5) A type of... Figure 4 The method shown is for the synthesis of (S)-2-(tert-butoxycarbonylamino)-3-(4-hydroxy-3-iodophenyl)propionic acid.

[0150] 6) The method of claim 5, further comprising: Figure 5 Synthesis of (S)-2-(tert-butoxycarbonylamino)-3-[3-iodo-4-(trimethylsilyloxy)phenyl]propionate trimethylsilyl ester.

[0151] 7) The method of claim 6, further comprising: Figure 6 shown 10 Synthesis of B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionic acid.

[0152] 8) The method of claim 7, further comprising: Figure 7 Synthesis of target material (4) to target material (5) as shown.

[0153] 9) A composition produced by a method of converting L-tyrosine into a target material, said method comprising the following synthesis:

[0154]

[0155] 10) The method of claim 9, wherein the resulting composition is... Figure 2 As shown in the image.

[0156] 11) A type of... Figure 10 The image shown is used to... 10 B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionic acid is converted to 10 A method for synthesizing methyl B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionate.

[0157] 12) The method of claim 11, further comprising: Figure 11 The synthesis of Boc-Tyr(3-B(OH)2, 4-OMe)-OMe to Boc-Tyr(3-B(OH)2, 4-OMe)-OH is shown.

[0158] 13) The method of claim 12, further comprising: Figure 12 Synthesis of target material (16) to target material (17) shown.

[0159] 14) A kit comprising the composition according to claim 1.

[0160] 15) A kit comprising the composition according to claim 2.

[0161] 16) A dosage unit comprising the composition according to claim 1.

[0162] 17) A dosage unit comprising the composition according to claim 2.

[0163] 18) The human unit form according to claim 16, wherein the human unit form is used for boron neutron capture therapy (BNCT).

[0164] 19) The human unit form according to claim 17, wherein the human unit form is used for boron neutron capture therapy (BNCT).

[0165] 20) The human unit form according to claim 16, wherein the human unit form is used for proton boron fusion therapy (PBFT).

[0166] 21) The human unit form according to claim 17, wherein the human unit form is used for proton boron fusion therapy (PBFT).

[0167] Example:

[0168] Various aspects of the present invention are further described and illustrated by the following examples, which are not intended to limit the scope of the invention.

[0169] Example 1: Preparation of TC220 hydrochloride.

[0170] Used to produce TC220 hydrochloride ( Figure 1 The overall synthesis scheme of ) in Figure 3 As shown in the figure, it is carried out using chemical methods.

[0171] Example 2: Preparation of (S)-2-(tert-butoxycarbonylamino)-3-(4-hydroxy-3-iodophenyl)propionic acid.

[0172] Use such as Figure 4 The starting material shown (chemical structure 1) was used to synthesize the following material using the following method.

[0173] In short, water (760 mL) and potassium hydroxide (63.1 g, 86.7%, 0.977 mol) were placed in a 3-L three-necked round-bottom flask equipped with a mechanical stirrer and thermometer. The mixture was stirred until the potassium hydroxide dissolved, and then 3-iodo-L-tyrosine (1) (150 g, 0.489 mol) was added. The reaction mixture was cooled to 9°C, and a solution of di-tert-butyl dicarbonate (107 g, 0.491 mmol) in THF (150 mL) was added. The cooling was then removed, and the mixture was stirred at room temperature for 18 hours.

[0174] The reaction mixture was then placed on a rotary evaporator and concentrated under reduced pressure at 45°C until it reached a weight of 700 g. The residue was then transferred to a 5-L three-necked round-bottom flask equipped with a mechanical stirrer and a pH meter. MTBE (700 mL) was added. The mixture was acidified to pH 3.1 by carefully adding citric acid hydrate in 20-g portions (totaling 240 g of citric acid hydrate). As observed, the mixture foamed.

[0175] Subsequently, heptane (350 mL) was added, and the upper organic layer was separated. The organic layer was separated, extracted twice with water (700 mL), and stirred with sodium sulfate (100 g) for 30 minutes. The suspension was filtered, and the filter cake was washed with MTBE (2 × 50 mL). The filtrate was concentrated under reduced pressure at 30 °C on a rotary evaporator in a 5-L flask until most of the organic solvent had evaporated. Heptane (200 mL) was added, and concentration was continued until the mixture foamed and filled most of the flask. More heptane (200 mL) was added, and the mixture was evaporated again until the organic solvent had stopped distilling. The residue was placed under vacuum and kept at room temperature for 3 days. This yielded the title compound (chemical structure 2) as a grayish-white solid foam (198 g, 99%). Confirmatory NMR was performed using standard methods, and the following results were obtained: 1 H NMR (500 MHz, CD3OD) 7.57 (d, J = 2.1 Hz, 1H), 7.06 (dd, J = 8.3, 2.1 Hz, 1H), 6.77 (d, J = 8.2 Hz, 1H), 4.28 (dd, J = 8.9, 5.0 Hz, 1H), 3.06 (dd, J = 14.0, 5.1 Hz, 1H), 2.79 (dd, J = 14.0, 9.0 Hz, 1H), 1.42 (s, 9H). (see Figure 4 ).

[0176] Example 3: (S)-2-(tert-butoxycarbonylamino)-3-[3-iodo-4-(trimethylsiloxy)phenyl]propionic acid Preparation of trimethylsilyl ester.

[0177] Use such as Figure 5 The starting material shown (chemical structure 2) was used to synthesize the following material using the following method.

[0178] In summary, N,N-dimethyltrimethylsilaneamine (117 mL, 0.732 mol) was charged into a 500-mL three-necked round-bottom flask equipped with a magnetic stirrer, temperature regulator, reflux condenser, and a nitrogen inlet above the condenser. Chemical structure 2 (60.0 g, 0.147 mol) was then added in portions over a nitrogen stream. Notably, bubbling and gas escape were observed. During the addition, the internal temperature of the mixture rose to 40 °C. The mixture was then heated under reflux at 94–96 °C for 18 hours. After heating, the temperature was lowered to 20 °C, and the mixture was then rapidly transferred to a 500-mL single-necked round-bottom flask. The excess reagent was then evaporated under reduced pressure on a rotary evaporator at 70 °C for 1 hour. After this interval, the flask was cooled back to 20 °C under vacuum and then filled with nitrogen. The title compound (chemical structure 3) (76.5 g) was obtained in 94% yield as a brown oil. (See...) Figure 5 ).

[0179] Example 4: 10 B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionic acid preparation.

[0180] Use such as Figure 6 The starting material shown (chemical structure 3) was used to synthesize the following material using the following method.

[0181] In short, chemical structure 3 (76.5 g, 139 mmol) was first dissolved in anhydrous THF (320 mL) under a nitrogen atmosphere. This solution was then transferred via a sleeve to a nitrogen-pre-charged 2-L three-necked flask equipped with a magnetic stirrer, a feeding funnel, a nitrogen inlet, and a thermometer. The contents of the flask were cooled to -25°C, and then a 2 M solution (175 mL) of isopropyl magnesium chloride in THF was added over a 30-minute period via the feeding funnel, maintaining an internal temperature of -27°C to -23°C. The funnel was rinsed with anhydrous THF (15 mL), which was also added to the mixture. The reaction was carried out at -27°C to -23°C for 1 hour. At this point, the mixture was cooled to -50°C, and then added over a 10-minute period via the feeding funnel at -55°C to -50°C. 10 Triisopropyl borate (65 mL, 283 mmol). After addition, the mixture is allowed to be gradually warmed to 7°C over a 2-hour period.

[0182] The reaction mixture was then further cooled to maintain an internal temperature below 10°C and quenched by adding a mixture of 2 M hydrochloric acid aqueous solution (170 mL) and water (140 mL). The pH of the reaction was adjusted to 3 using 1 M sodium hydroxide aqueous solution. The organic phase was separated and transferred to a 500-mL three-necked flask equipped with a magnetic stirrer, pH meter, feeding funnel, and nitrogen inlet. Then, 1 M sodium hydroxide aqueous solution (approximately 175 mL) was added under nitrogen until the pH of the mixture reached 6.85. The resulting mixture was then concentrated using a rotary evaporator under reduced pressure (without heating) for 4 hours until a residue weighing 270 g was obtained.

[0183] The mixture was then extracted twice under nitrogen using MTBE (280 mL, then 250 mL). The aqueous layer was separated and transferred to a 1-L three-necked flask equipped with a magnetic stirrer, pH meter, feeding funnel, and nitrogen inlet. MTBE (300 mL) was added to the aqueous layer, followed by dropwise addition of concentrated 37% hydrochloric acid (approximately 16 g) under nitrogen to achieve a pH of 3.1. The organic layer was rapidly separated and stirred for 10 minutes under nitrogen atmosphere with magnesium sulfate (26 g). The suspension was rapidly filtered, and the filter cake was washed with MTBE (2 × 25 mL). The filtrate was evaporated under reduced pressure (without heating) using a rotary evaporator and then maintained under vacuum for 18 hours to produce crude chemical structure 4 (39.5 g) as a grayish-white foam.

[0184] Finally, the foam was stirred with ethyl acetate (550 mL) under nitrogen atmosphere for 4 hours. The resulting suspension was then filtered through a 10-µm pad filter, and the filter cake was dried under vacuum for 4 days. The resulting solid was retreated with ethyl acetate as outlined above to yield a pure title compound (chemical structure 4) as a grayish-white solid (20.1 g, 45% yield). Confirmatory NMR was performed using standard methods, and the following results were obtained: 1 H NMR (500 MHz, CD3OD) 7.65-7.0 (br m, 2H), 6.77 (d, J = 8.3 Hz, 1H), 4.27 (dd, J = 8.5, 5.0 Hz, 1H), 3.05 (dd, J = 13.8, 5.1 Hz, 1H), 2.83 (dd, J = 13.9, 8.7 Hz, 1H), 1.38 (s,9H). (see Figure 6 ).

[0185] Example 5: 10 Preparation of B-TC220 hydrochloride.

[0186] Use such as Figure 7The starting material shown (chemical structure 4) was used to synthesize the following material using the following method.

[0187] In short, dioxane (250 mL) and a 4 M solution of hydrogen chloride in dioxane (250 mL) were added to a 1-L three-necked flask equipped with a magnetic stirrer and a nitrogen inlet. Chemical structure 4 (24.7 g, 76.2 mmol) was added in portions under a nitrogen atmosphere. The mixture was then stirred at ambient temperature for 6 hours and filtered through a 10 µm pad filter. The resulting filter cake was dried overnight under vacuum at room temperature.

[0188] The dried filter cake was then dissolved in water (140 mL), and the filtrate was clarified through a 2.5 µm filter. It was subsequently lyophilized to obtain a white solid. 10 B-TC220 hydrochloride (chemical structure 5) (see also...) Figure 1 The product, with a yield of 86% (17.0 g), was subjected to confirmatory NMR using standard methods, and the following results were obtained: 1 H NMR (500 MHz, D2O) 7.36 (d, J = 2.4 Hz, 1H), 7.16 (dd, J = 8.4, 2.4 Hz, 1H), 6.77 (d, J = 8.3Hz, 1H), 4.13 (dd, J = 7.7, 5.5 Hz, 1H), 3.15 (dd, J = 14.8, 5.5 Hz, 1H),3.03 (dd, J = 14.8, 7.7 Hz, 1H). (see Figure 7 ).

[0189] Example 6: 10 Purity analysis of B-TC220 hydrochloride.

[0190] Purity analysis of TC220 was performed using standard methods in the art. In short, UPLC / MS was performed using a C18 column with a gradient of 2% to 20% B. Solvent conditions were 0.1% formic acid (A) / water and 0.1% formic acid (B) / acetonitrile, at a flow rate of 0.500 mL / min. Maximum absorbance was observed at 225 nm.

[0191] Example 7: Synthetic scheme for converting TC220 hydrochloride to TC221.

[0192] Used to treat TC220 hydrochloride ( Figure 1 ) transformed into TC221 ( Figure 2 The overall synthesis scheme of ) in Figure 9 As shown in the figure, it is carried out using chemical methods.

[0193] Example 8: Used to... 10 B-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl] propionic acid converted to 10 Methyl β-(S)-2-(tert-butoxycarbonylamino)-3-[3-(dihydroxyboryl)-4-hydroxyphenyl]propionate Synthesis scheme.

[0194] Use such as Figure 9 The starting material shown (chemical structure 4) was used to synthesize the following material using the following method.

[0195] In short, K₂CO₃ (7.4 g, 53.51 mmol) and Me₂SO₄ (2.5 g, 44.6 mmol) were added to a solution of chemical structure 4 (5.0 g, 17.84 mmol) in acetone (50 mL) at room temperature. The reaction mixture was stirred at the same temperature for 12 hours. The reaction mixture was filtered through a diatomaceous earth mat and washed with acetone (50 mL). The acetone layer was concentrated under vacuum to obtain chemical structure 15, which was a grayish-white viscous material. (See also...) Figure 10 ).

[0196] Example 9: Preparation of (S)-2-(tert-butoxycarbonylamino)-3-(4-methoxy-3-boronic acid)propionic acid.

[0197] The synthesis of N-Boc-Tyr(3-B(OH)2, 4-OMe)-OMe to N-Boc-Tyr(3-B(OH)2, 4-OMe)-OH was carried out in Figure 11 The modification is shown and performed using standard methods. See Gao et al., Arkivoc (2009) (vii) pp. 72-78. This modification does not utilize a procedure for demethylation using BBr3, and... Figure 9 As shown in the diagram, this is to maintain the dimethyl ether.

[0198] Example 10: Preparation of (S)-2-(amino)-3-(4-methoxy-3-boronic acid)propionic acid, i.e., TC221.

[0199] The synthesis of N-Boc-Tyr(3-B(OH)2, 4-OMe)-OH to TC221 was carried out in Figure 12 The following describes and utilizes further synthetic modifications within the scope of this disclosure. In short, a solution of 4 M hydrochloric acid in dioxane was added to a flask containing 1.0 g of N-Boc-Tyr(3-B(OH)2, 4-Ome)-OH. After one (1) hour, no Boc-protected starting material was observed. Volatile solvents and acids were removed under reduced pressure, and the target material was purified by preparative LC using the described protocol. In short, a C18 preparative column with dimensions of 30 mm × 100 mm was used. The flow rate was set to 50 mL / min. The gradient used was expressed as follows: 0% acetonitrile to 20% acetonitrile / water over 20 minutes.

[0200] The resulting TC221 has Figure 2 The structure shown.

[0201] Example 11: Human clinical trial of treating human cancer using TC220 and TC221.

[0202] TC220 and / or TC221 are synthesized according to the present invention, and specifically accumulate in tumor cells for the treatment of certain tumors and other immune disorders and / or other diseases. Combining each of these indications, two clinical approaches have been successfully obtained.

[0203] I.) adjunctive therapy In adjuvant therapy, patients are treated with TC220 and / or TC221 in combination with chemotherapeutic agents, drugs, or biologics. Primary cancer targets are treated with the addition of TC220 and / or TC221 under a standard regimen, followed by irradiation. The regimen design addresses the efficacy evaluated in examples such as those described below, including but not limited to reduction in tumor mass of primary or metastatic lesions, progression-free survival, increased overall survival, improved patient health, disease stabilization, and the ability to reduce the usual doses of standard chemotherapy and other biologics. These dose reductions allow for additional and / or prolonged therapy by reducing dose-related toxicities of chemotherapeutic agents or biologics.

[0204] II.) Monotherapy This involves combining the use of TC220 and / or TC221 as monotherapy for tumors, administering the TC220 and / or TC221 to patients in the absence of chemotherapeutic agents, drugs, or biologics. In one embodiment, monotherapy is performed clinically in patients with advanced cancer and extensive metastatic disease. The protocol design addresses efficacy assessed as demonstrated in the following examples, including but not limited to reduction in tumor mass of primary or metastatic lesions, progression-free survival, increased overall survival, improved patient health, disease stabilization, and the ability to reduce the usual doses of standard chemotherapy and other biologics.

[0205] dose

[0206] Dosing regimens can be adjusted to provide the optimal desired response. For example, a single injection of TC220 and / or TC221 can be administered, several fractionated doses can be administered over time, or the dose can be reduced or increased proportionally as indicated by the urgency of the treatment situation. As used herein, “dosage unit form” refers to a physically discrete unit suitable as a unit dose to a mammalian subject to be treated; each unit contains a predetermined amount of the active compound, said predetermined amount being calculated to produce the desired therapeutic effect associated with the desired drug delivery. The specifications of the dosage unit form of the present invention are determined by and directly depend on: (a) the unique characteristics of TC220 and / or TC221, the various mechanics of the irradiation apparatus (reactor), and the specific therapeutic or preventative effect to be achieved; and (b) the inherent limitations in the art of combining such compounds for the individual’s sensitivity to treatment.

[0207] Clinical Development Program (CDP)

[0208] CDP follows and develops treatments for cancer and / or immune disorders using TC220 and / or TC221 disclosed herein, followed by irradiation with neutron capture therapy in combination with adjuvant or monotherapy. Trials initially demonstrated safety and subsequently confirmed efficacy with repeated doses. The trials were open-label, comparing standard chemotherapy with standard therapy plus TC220 and / or TC221 followed by irradiation with boron neutron capture therapy. As will be appreciated, one non-limiting criterion for patient inclusion may be the concentration of TC220 and / or TC221 in the tumor, determined by standard detection methods known in the art.

[0209] This invention is not limited to the embodiments disclosed herein, which are intended as simple illustrations of various aspects of the invention, and any functionally equivalent embodiments are within the scope of the invention. Various modifications to the models, methods, and lifecycle methodologies of the invention, in addition to those described herein, will become apparent to those skilled in the art from the foregoing description and teachings and are similarly intended to fall within the scope of the invention. These modifications or other embodiments may be practiced without departing from the true scope and spirit of the invention.

[0210] Table I. Naturally occurring amino acids .

[0211]

[0212]

Claims

1. A method of... L -A composition produced by a method for converting tyrosine into a target material, said method comprising the following synthesis: 。 2. A method of... L -A composition produced by a method for converting tyrosine into a target material, said method comprising the following synthesis: 。 3. The method of claim 1, wherein the resulting composition is designated TC220 and is shown in FIG1.

4. The method of claim 2, wherein the resulting composition is designated TC221 and is shown in FIG2.

5. A kit comprising a composition produced by the method according to claim 1.

6. A kit comprising a composition produced by the method according to claim 2.

7. A dosage unit comprising a composition produced by the method according to claim 1.

8. A dosage unit comprising the composition produced by the method according to claim 2.

9. The dosage unit form according to claim 7, wherein the dosage unit form is for boron neutron capture therapy (BNCT).

10. The dosage unit form according to claim 8, wherein the dosage unit form is for boron neutron capture therapy (BNCT).

11. The dosage unit form according to claim 7, wherein the dosage unit form is for proton boron fusion therapy (PBFT).

12. The dosage unit form according to claim 8, wherein the dosage unit form is for proton boron fusion therapy (PBFT).

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