Processes for producing reduced hydroxy-substituted isoflavone, processes for producing pterocarpan, pterocarpan compound, reduced hydroxy-substituted isoflavone, hydroxy-substituted isoflavone, liposome, process for producing the liposome, pharmaceutical composition, use of the pterocarpan compound, the liposome and the pharmaceutical composition, and method of treatment

Abstract

The invention relates to a synthetic route for pterocarpan compounds, to a pterocarpan nanoencapsulated in a liposome or in an immunoliposome functionalized with a monoclonal antibody, to a pharmaceutical composition comprising the liposomes or immunoliposomes, and to use of the pharmaceutical formulation for manufacturing a medicament for the treatment of cancer.

Description

PRODUCTION PROCESSES FOR HYDROXY-SUBSTITUTED REDUCED ISOFLAVONE, PRODUCTION PROCESSES FOR PTEROCARPANE, PTEROCARPANE COMPOUND, HYDROXY-SUBSTITUTED REDUCED ISOFLAVONE, HYDROXY-SUBSTITUTED ISOFLAVONE, LIPOSOME, PRODUCTION PROCESS FOR LIPOSOME, PHARMACEUTICAL COMPOSITION, USE OF PTEROCARPANE COMPOUND, LIPOSOME AND PHARMACEUTICAL COMPOSITION AND TREATMENT METHOD Technical Field

[0001] The invention relates to a synthetic route for pterocarpan compounds, to a pterocarpan nanoencapsulated in liposomes or immunoliposomes functionalized with monoclonal antibodies, to a pharmaceutical composition comprising the liposomes or immunoliposomes, and to the use of the pharmaceutical formulation to manufacture a drug for the treatment of cancer. The present invention falls within the fields of Nanotechnology, Chemistry, Medicine and Biology. Fundamentals of the Invention

[0002] Historically, nature has been a rich source of medicinal compounds. Plants, microorganisms, and animals produce secondary metabolites that serve as defense mechanisms and have therapeutic properties. Natural products remain essential in modern medicine, with 32% of approved drugs between 1981 and 2019 being derived from such sources. Plants are a significant source of biologically active compounds such as terpenes, alkaloids, and flavonoids.

[0003] Among the natural compounds highlighted for the treatment of diseases, paclitaxel from Taxus brevefolia is widely used in the treatment of breast cancer. Alkaloids from vinca, such as vinblastine and vincristine, isolated from Catharanthus roseus, are used to treat lymphomas. Microorganisms have also contributed to the discovery of antitumor compounds, such as doxorubicin from the bacterium Streptomyces peucetius, used against solid and hematological tumors.

[0004] Isoflavonoids have demonstrated significant antitumor potential. Compounds such as genistein, daidzein, and pterocarpans exhibit activity against various types of cancer, acting through mechanisms such as inhibition of DNA topoisomerase and tyrosine kinases.

[0005] A subclass of isoflavonoids, the pterocarpans, are produced by various plants and have diverse biological activities, including antifungal, antiviral, antibacterial, anti-inflammatory, antiosteoporotic, and anticancer actions. Medicarpin, isolated from the plant *Medicago sativa*, has shown efficacy against leukemic cells, including those resistant to multiple drugs, inducing apoptosis via the mitochondrial pathway. Another relevant pterocarpan is (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpan ((+)-PTC), isolated from *Platymiscium floribundum*, which has demonstrated high selective cytotoxicity against several cancer cell lines. Studies indicate that this molecule is metabolized by hepatic cytochrome P450 enzymes.

[0006] Despite the promising biological activities of pterocarpans, obtaining them from natural sources presents significant challenges, such as low yield and variability in metabolite concentration. To overcome these limitations and enable industrial-scale production, the search for efficient synthetic routes is crucial. Such routes should not only provide high yield but also, ideally, be serenantioselective, allowing the synthesis of the isomer with the highest biological activity in a pure and consistent manner, and capable of being scaled up to an industrial level. Synthetic production offers a controlled and scalable solution to fully exploit the therapeutic potential of these compounds.

[0007] Furthermore, there is a need to improve the pharmacokinetic parameters of pterocarpan compounds. Therefore, knowing that several antitumor drugs, such as paclitaxel, demonstrate high efficacy against cancers but face clinical limitations due to low accumulation in target tissue and adverse effects, such as neurotoxicity leading to sensory axonal neuropathy in approximately 30% of patients, it is essential to develop strategies to optimize their use. An effective approach is the use of nanoparticles as drug delivery systems, since they offer greater stability, biocompatibility, permeability, retention, and... Targeting the tumor region improves pharmacokinetic parameters and reduces side effects and drug resistance. Different types of nanoparticles are being studied for drug encapsulation, including polymeric micelles, liposomes, and functionalized nanoparticles. Liposomes allow the encapsulation of hydrophobic and hydrophilic drugs due to their lipid bilayer, being biocompatible and biodegradable, improving the biodistribution and stability of the drugs.

[0008] The first liposome approved for cancer treatment was DOXIL, in 1995, which encapsulates doxorubicin and includes PEG in its formulation; PEG increases the half-life of liposomal formulations, reducing the cardiotoxicity of doxorubicin and improving pharmacokinetic parameters without losing efficacy in the treatment of Kaposi's lymphoma and ovarian cancer. Therefore, liposomal formulations facilitate drug delivery through passive targeting, exploiting the specific characteristics of tumor tissues, such as high cell proliferation, angiogenesis, and increased vascular permeability, as well as reduced lymphatic drainage; these factors promote greater retention and accumulation of liposomes in the tumor microenvironment through the EPR ("Enhanced Permeability and Retention") phenomenon.

[0009] However, passive targeting has limitations, such as non-specific drug distribution and variations in vascular permeability between different tumors. As an effective strategy to overcome these limitations, active targeting functionalizes the surface of liposomes with specific ligands, such as monoclonal antibodies, peptides, or proteins, directing them to specific tumor cells and increasing treatment efficacy while reducing side effects.

[0010] In addition to conventional drugs, research is being conducted on nanoformulations with natural products, which in some cases have low solubility and bioavailability. For example, quercetin, a flavonoid with antitumor activity, faces clinical challenges due to its low bioavailability and high hydrophobicity; to improve these parameters, a DSPE-PEG2000 nanomicelle was developed to encapsulate quercetin, resulting in better antitumor activity and greater accumulation in the tumor compared to the free compound.

[0011] In the search for the state of the art in scientific and patent literature, the following documents were found that address the topic:

[0012] Patent document BR132014024531-5 discloses the use of pterocarpan in a cytomodulatory composition. Patent document WO2015034436 discloses the specific use of a flavonoid polymer conjugated with compounds containing hyaluronic acid for the treatment of cancer.

[0013] Thus, based on the literature reviewed, there is a need not only to develop effective and enantioselective synthetic routes for pterocarpan compounds, but also to develop efficient drugs for transporting pterocarpans for cancer treatment. The present invention aims to meet these needs through new effective synthesis routes for obtaining pterocarpan compounds and from pterocarpan nanoencapsulates using liposomes and functionalized immunoliposomes for cancer treatment. As will be seen from the description and examples, the present invention offers advantages in increasing the yield of pterocarpan compounds (PTCs) and / or in the possibility of scaling up production, as well as being able to use them in pharmaceutical nanoformulations for greater efficiency in transporting targeted drugs for cancer treatment, in addition to reducing the adverse effects of the pterocarpan compound. Summary of the Invention

[0014] In a first aspect, the present invention relates to a process for producing hydroxy-substituted reduced isoflavone of formula (II): comprising the asymmetric hydrogen transfer step by contact of hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya Ruthenium (S,S) complex in the presence of a hydrogen donor, in which the process is enantioselective for the reduced hydroxy-substituted isoflavone (S,S) of formula (Ha): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

[0015] In a second aspect, the present invention relates to a process for producing hydroxy-substituted reduced isoflavones of formula (Ha): comprising the asymmetric hydrogen transfer step by contact of hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya complex of Ruthenium (S,S) in the presence of a hydrogen donor, wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy.

[0016] In a third aspect, the present invention relates to a process for producing pterocarpane of formula (I): understanding the acid cyclization step of the reduced hydroxy substituted isoflavone of formula (II): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

[0017] In a fourth aspect, the present invention relates to a process for producing pterocarpane of formula (la): understanding the acid cyclization step of the reduced hydroxy-substituted isoflavone with the formula (Ha): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy.

[0018] In a fifth aspect, the present invention relates to a process for producing pterocarpane of formula (I): understanding the reductive cyclization step of the hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya ruthenium (S,S) complex in the presence of a hydrogen donor, in which the process is enantioselective for pterocarpane (S,S) of formula (1a): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

[0019] In a sixth aspect, the present invention relates to a process for producing pterocarpane of formula (1a): understanding the enantioselective reductive cyclization step of the hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya complex of Ruthenium (S,S) in the presence of a hydrogen donor, wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy.

[0020] In a seventh aspect, the present invention relates to a pterocarpane compound of formula (I): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy.

[0021] In an eighth aspect, the present invention relates to a hydroxy-substituted reduced isoflavone of formula (II): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy or haloalkoxy.

[0022] In a ninth aspect, the present invention relates to a hydroxy-substituted isoflavone of formula (III): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy or C1-C6 haloalkoxy.

[0023] In a tenth aspect, the present invention relates to a liposome comprising lipids and a pterocarpane compound.

[0024] In an eleventh aspect, the present invention relates to a liposome production process described herein comprising the steps of: (a) dilution of the support phase material in a volatile organic solvent, (b) formation of a lipid film, (c) evaporation of the volatile organic solvent, (d) solubilization of the lipid film in an aqueous phase, (e) fragmentation to obtain unilamellar nanoparticles.

[0025] In a twelfth aspect, the present invention relates to a pharmaceutical composition comprising the pterocarpan compound of formula (I) described herein, or the liposome described herein.

[0026] In a thirteenth aspect, the present invention relates to the use of the pterocarpan compound described herein, the liposome described herein, or the pharmaceutical composition described herein in the manufacture of a medicament for treating cancer.

[0027] In a fourteenth aspect, the present invention relates to a treatment method comprising administering the pterocarpan compound of formula (I) described herein, the liposome described herein, or the pharmaceutical composition described herein to an individual with cancer. Brief Description of the Figures

[0028] Figure 1 shows the nuclear magnetic resonance (NMR) spectrum of 1 H of the intermediate compound (1-(2-hydroxy-4,5-dimethoxypheni)ethan-1-one), obtained in the first step of Example 1.

[0029] Figure 2 shows the nuclear magnetic resonance (NMR) spectrum of 1 H of the intermediate compound 3-iodo-6,7-dimethoxy-4H-chromen-4-one, obtained in the second step of Example 1.

[0030] Figure 3 shows the nuclear magnetic resonance (NMR) spectrum of 1 H of the intermediate compound (3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one, obtained in the third step of Example 1.

[0031] Figure 4 shows the nuclear magnetic resonance (NMR) spectrum of 13 C of the intermediate compound (3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one, obtained in the third step of Example 1.

[0032] Figure 5 shows the nuclear magnetic resonance (NMR) spectrum of 1 H of the intermediate compound in 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4 / - / -chromen-4-one, obtained in the fourth step of Example 1.

[0033] Figure 6 shows the nuclear magnetic resonance (NMR) spectrum of 1 H of the intermediate compound (+)-(6aS,11aS)-2,3,9-trimethoxy-6a,11a-dihydro-6H-benzofuro[3,2-c]chromene, obtained in the fifth step of Example 1.

[0034] Figure 7 shows the nuclear magnetic resonance (NMR) spectrum of 1 H of the compound (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane, obtained in the sixth step of Example 1.

[0035] Figure 8 shows the high-performance chromatography of the compound (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane and the compound (-)-(6aS,11aS)-2,3,9-trimethoxypterocarpane, followed by the table corresponding to the data obtained.

[0036] Figure 9 shows the chromatography spectrum of (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane and the corresponding table of data obtained.

[0037] Figure 10 shows the size data of the nanoformulation encapsulated with the lead (+)-PTC compound of formulation 3, obtained using Dynamic Light Scattering (DLS) equipment.

[0038] Figure 11 shows the zeta potential data of the nanoformulation encapsulated with the lead (+)-PTC compound of formulation 3, obtained using Dynamic Light Scattering (DLS) equipment.

[0039] Figure 12 shows the stability of the size, polydispersity index (PDI), and zeta potential parameters of lyophilized liposome samples on different days.

[0040] Figure 13 shows the steps of immunoliposome functionalization with the monoclonal antibody. In A, the liposome is produced; in B, a thiol group is added to the monoclonal antibody; and in C, the liposome is incubated with the monoclonal antibody: immunoliposome.

[0041] Figure 14 shows the chromatogram of the formulation functionalized with anti-EGFR antibody.

[0042] Figure 15 shows the percentage of cellular internalization of the liposome and purified immunoliposome at different times.

[0043] Figure 16 shows the absorbance comparison in the 450 nm range, representing the ability of the Negative Control, Liposome, Total Immunoliposome, Purified Immunoliposome, and Cetuximab samples to bind to the EGF receptor.

[0044] Figure 17 shows the cell growth inhibition curve of the DU145 cell line after 72 hours of treatment with different samples. Definition of radicals Markush

[0045] The term "Ci-C6 alkyl" refers to a saturated, monovalent hydrocarbon radical containing 1 to 6 carbon atoms. The term encompasses both linear and branched chains. Examples of Ci-C6 alkyl groups include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, and all pentyl and hexyl isomers.

[0046] The term "Ci-Ce haloalkyl" refers to a saturated, monovalent hydrocarbon radical containing 1 to 6 carbon atoms, where one or more hydrogen atoms have been replaced by one or more halogen atoms. The halogen can be fluorine, chlorine, bromine, or iodine. The term encompasses both linear and branched chains. Examples of haloalkyl groups The O-C groups include, but are not limited to, difluoromethyl, trifluoromethyl, dichloromethyl, chloromethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, 2-bromo-1-methylpropyl, and 1-chloro-2-methylpropyl.

[0047] The term "Ci-Ce alkoxy" refers to a monovalent group with the formula R'-O-, where R' is a linear or branched alkyl group containing 1 to 6 carbon atoms. The term encompasses all possible structural isomers within this carbon range. Examples of Ci-Ce alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, and all pentoxy and hexoxy isomers.

[0048] The term "C1-C6 haloalkoxy" refers to a monovalent group with the formula R"-O-, where R is a linear or branched alkyl group containing 1 to 6 carbon atoms, in which one or more hydrogen atoms have been replaced by one or more halogen atoms. The halogen can be fluorine, chlorine, bromine, or iodine. The term encompasses all possible structural isomers within this carbon range. Examples of C1-C6 haloalkoxy groups include, but are not limited to, difluoromethoxy, trifluoromethoxy, 2-chloroethoxy, 2,2,2-trifluoroethoxy, and 3-bromopropoxy.

[0049] The term “q 6 -Ar” denotes an arene group of hapticity 6, where 'Ar' refers to an aromatic ligand that is coordinated to a metal atom through all six carbon atoms in the aromatic ring. The q notation 6This indicates that all six carbon atoms in the aromatic ring participate in the bonding with the metal, using the ring's pi electron system to form a stable complex. The term "arene" refers to a radical or a cyclic aromatic hydrocarbon group. In the context of the present invention, the aromatic ring of the arene is preferably a six-membered monocyclic group. The arene may be unsubstituted or optionally substituted with C1-C1 alkyl or C1-C1 alkoxy.

[0050] The term "optionally substituted" in relation to the arene ligand refers to an aromatic ring that may be unsubstituted or substituted at one, two, three, four, five, or even six carbon positions. Each of these substituents, when present, is independently selected from the list of radical groups defined here.

[0051] The term "arylsulfonyl" refers to a monovalent radical of formula Arl-SC-, where Ari is an aryl radical. The term "aryl" in the context of the present invention encompasses radicals derived from aromatic rings, such as benzene or naphthalene, which may be monocyclic or polycyclic, and which may optionally be substituted with one or more groups such as C1-C6 alkyl, C1-C6 alkoxy, or halogen. Examples of arylsulfonyl groups include, but are not limited to: phenylsulfonyl (Fs), p-toluenesulfonyl (Ts), and naphthalenesulfonyl (Ns). Detailed Description Enantioselective Production Process of Hydroxy-Substituted Reduced Isoflavone

[0052] In a first aspect, the present invention relates to a process for producing hydroxy-substituted reduced isoflavone of formula (II): comprising the asymmetric hydrogen transfer step by contact of hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya Ruthenium (S,S) complex in the presence of a hydrogen donor, wherein the process is enantioselective for the reduced hydroxy-substituted isoflavone (S,S) of formula (Ha): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

[0053] The term "asymmetric hydrogen transfer" refers to a process in which a hydrogen donor transfers a hydrogen atom to a prochiral molecule. What makes the process asymmetric is the use of the chiral catalyst, the Ruthenium (S,S) Noyori-lkariya complex, which directs the reaction to preferentially produce the (S,S) enantiomer rather than a racemic mixture.

[0054] Among the advantages of using hydroxy-substituted isoflavones as a reagent is the fact that the C=O bond of the hydroxy-substituted isoflavone of formula (III) is more easily reduced in the asymmetric hydrogen transfer reaction in substrates capable of forming intramolecular hydrogen bonds, thus requiring lower catalytic charges (<1 mol%).

[0055] In the context of the present invention, the term "enantioselective process" refers to a chemical synthesis capable of directing the reaction towards the preferential formation of one of the enantiomers of the product to the detriment of the other enantiomers. Unlike conventional processes that result in a racemic mixture, the present process is notably advantageous in providing the desired enantiomer with a high enantiomeric excess. The ability to preferentially synthesize a single enantiomer in a controlled and efficient manner is of paramount importance in the pharmaceutical field, where the biological activity and safety of a compound are intrinsically linked to its chirality, conferring greater efficacy and a more precise therapeutic profile to the final product. In one or more embodiments, the enantiomeric excess of the (S,S) isomer is at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98% or at least 99% on a molar basis.

[0056] In one or more embodiments, the C1-C4 alkyl radicals are selected independently from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl. In one or more embodiments, the C1-C4 alkyl radicals are selected independently from C1-C4 alkyl, preferably those listed here.

[0057] In one or more embodiments, the C1-C4 haloalkyl radicals are selected independently from fluoromethyl, chloromethyl, bromomethyl, iodomethyl, trifluoromethyl, 2-fluoroethyl, 2-chloroethyl, 2,2,2-trifluoroethyl, 3-chloropropyl, and 4-chlorobutyl. In one or more embodiments, the C1-C4 haloalkyl radicals are selected independently from C1-C4 haloalkyl, preferably those listed herein.

[0058] In one or more embodiments, the C1-C4 alkoxy radicals are selected independently from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy. In one or more embodiments, the C1-C4 alkoxy radicals are selected independently from methoxy, ethoxy, n-propoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy. In one or more embodiments, the C1-C4 alkoxy radicals are selected independently from methoxy.

[0059] In one or more embodiments, the C1-C4 haloalkoxy radicals are independently selected from fluoromethoxy, difluoromethoxy, trifluoromethoxy, 2-chloroethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, bromomethoxy, 3-chloropropoxy, 2-chloroisopropoxy, and 2,2-dichloroethoxy. In one or more embodiments, the C1-C4 haloalkoxy radicals are independently selected from C1-C4 haloalkoxy, preferably those listed here. In one or more embodiments, the C1-C4 haloalkoxy radicals are difluoromethoxy.

[0060] In one or more embodiments, R1, R2, and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen, hydroxyl, Ci-Ce alkoxy, or Ci-Ce haloalkoxy; and R5, R6, R7, and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

[0061] Preferred embodiments of radicals R1 to R8 are presented in Table 1 below. Table 1

[0062] Preferably, in the embodiments presented in Table 1, the C1-C1 alkoxy radicals are independently selected from methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy. More preferably, the C1-C1 alkoxy radicals in Table 1 are methoxy. Also in Table 1, preferably the C1-C1 haloalkoxy radicals are independently selected from fluoromethoxy, difluoromethoxy, trifluoromethoxy, 2-chloroethoxy, 2-fluoroethoxy, 2,2,2-trifluoroethoxy, bromomethoxy, 3-chloropropoxy, 2-chloroisopropoxy, and 2,2-dichloroethoxy. More preferably, the C1-C1 haloalkoxy radicals in Table 1 are difluoromethoxy. In an even more preferred embodiment, the C1-C1 alkoxy radicals of Table 1 are methoxy and the C1-C1 haloalkoxy radicals of Table 1 are difluoromethoxy.The most preferred embodiments are listed in Table 2, where each combination is assigned a specific number for compounds II and III, and OME represents a methoxy radical. Table 2

[0063] In one or more embodiments, R1, R2, R3, R4, R5, R6, R7, and R8 are selected such that compound II is I1.1, II.2, II.3, II.4, II.5, II.7, II.12, II.17, II.22, II.27, II.32, II.37, II.42, and / or II.47, obtained from the corresponding compounds III.

[0064] The asymmetric hydrogen transfer step occurs in the presence of a hydrogen donor. In one or more embodiments, the hydrogen donor is selected from carboxylic acid salts, carboxylic acids, amines, or a combination thereof. Preferred carboxylic acid salts according to the present invention are C1-C10 carboxylic acid salts. The term "C1-C10 carboxylic acids" refers to organic compounds having one to six carbon atoms in their structure, containing a single carboxyl group (-COOH). These compounds may be saturated or unsaturated, linear or branched chain. Non-limiting examples include formic acid, acetic acid, and propionic acid. Preferably, the carboxylic acid salts are selected from alkali metal C1-C10 carboxylic acid salts, preferably alkali metal formates. The alkali metals are preferably selected from lithium, sodium, or potassium.In one or more embodiments, the alkali metal C1-C6 carboxylic acid salts are lithium formate, sodium formate, or potassium formate, preferably sodium formate. Carboxylic acids may preferably be selected from C1-C6 carboxylic acids, preferably formic acid. Amines may be selected from C1-C6 trialkylamines, wherein the term “C1-C6 trialkylamines” refers to tertiary amines having three alkyl groups, which may be linear or branched, and containing from one to six carbon atoms in total between the three groups. The alkyl groups may be the same or different. Preferred amines include trimethylamine and triethylamine. In one or more embodiments, carboxylic acids and amines are used in combination, wherein the amine acts as a co-donor together with the carboxylic acid. Preferably, the combination used is triethylamine with formic acid.

[0065] The asymmetric hydrogen transfer step occurs in the presence of a Noyori-lkariya ruthenium (S,S) complex. The term "Noyori-lkariya ruthenium (S,S) complex" refers to a type of catalytic organometallic complex containing a transition metal, specifically ruthenium, coordinated to a chiral diamine ligand structure and an arene ligand structure. The "(S,S)" annotation specifies that the two chiral carbon centers in the diamine ligand have the absolute S spatial configuration, as per the Cahn-Ingold-Prelog nomenclature. In one or more embodiments, the Noyori-lkariya ruthenium (S,S) complex is a Noyori-lkariya ruthenium (S,S) complex of formula (VII): where M is ruthenium; q6-Ar is an arene ligand of hapticity 6 linked to M selected from optionally substituted arene with Ci-Ce alkyl or Ci-Ce alkoxy; Ph is phenyl; X is halogen; and R13 is arylsulfonyl.

[0066] In one or more embodiments, the optionally substituted arene is selected from the C1-C1 alkyl substituted arene, which may be substituted at one, two, three, four, five, or six carbons of the aromatic ring. Preferably, the C1-C1 alkyl substituted arene is selected from 1-isopropyl-4-methylbenzene (p-cymene), 1,3,5-trimethylbenzene (mesitylene), hexamethylbenzene, and methylbenzene. In one or more embodiments, the arene is unsubstituted (benzene). In one or more embodiments, the halogen X is selected from fluorine, chlorine, bromine, or iodine, preferably chlorine. In one or more embodiments, the arylsulfonyl is selected from phenylsulfonyl (Fs), p-toluenesulfonyl (Ts), and naphthalenesulfonyl (Ns).

[0067] The rate of the asymmetric hydrogen transfer reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the process be conducted over a period of 2 to 24 hours. In one or more embodiments, the asymmetric hydrogen transfer reaction is carried out in a temperature range of 25 °C to 45 °C.

[0068] Due to the presence of the hydroxyl group in the ortho (2') position on the phenyl ring of the hydroxy-substituted reduced isoflavone of formula III, the reduction of the molecule is facilitated, allowing the use of catalytic charges of the Noyori-lkariya complex. Ruthenium levels equal to or less than 2 mol%, more preferably equal to or less than 1 mol%, and even more preferably less than 1 mol%.

[0069] The hydroxy substituted isoflavone of formula (III) used as a reagent in the asymmetric hydrogen transfer reaction can be produced from the selective dealkylation of the alkyloxy substituted isoflavone of formula (IV): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C6 alkoxy, or C1-C1 haloalkoxy, and R9 is selected from C1-C1 alkyl.

[0070] The term "selective dealkylation" refers to a chemical process that removes one or more alkyl groups from a molecule in a controlled manner, without affecting other functional groups or reactive centers present in the same structure. In the present invention, the alkyl group removed in selective dealkylation is the C1-C1 alkyl group of the R9 radical.

[0071] In one or more embodiments, R9 is selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, and n-hexyl. In one or more embodiments, R9 is selected from C1-C4 alkyl groups, preferably those listed here. More preferably, R9 is methyl, and selective dealkylation may be termed selective demethylation.

[0072] All embodiments of R1, R2, R3, R4, R5, R6, R7, and R8 previously realized for compounds (II) and (III) also apply to compound IV, including the preferred embodiments of Tables 1 and 2. For compound IV, Table 1 can be supplemented by R9 being a C1-C6 alkyl, combining all listed embodiments. More preferably, R9 is methyl, and the preferred compounds IV include radical combinations of Table 2 and R9 being methyl, and are designated IV.1 to IV.50, respectively, following the same numbering proposed for compounds II and III. In one or more embodiments, R1, R2, R3, R4, R5, R6, R7, R8, and R9 are selected such that compound IV is IV.1, IV.2, IV.3, IV.4, IV.5, IV.7, IV.12, IV.17, IV.22, IV.27, IV.32, IV.37, IV.42, and / or IV.47.

[0073] In one or more embodiments, selective dealkylation occurs in the presence of a Lewis acid. Lewis acids used in selective dealkylation are preferably boron trichloride (BCh) or aluminum halides, more preferably aluminum chloride (AlCh), aluminum bromide (AIBrs), aluminum iodide (AIH), aluminum fluoride (AIF3), and ethyl aluminum sesquichloride (EtsAPCI). 3 ), wherein boron trichloride and aluminum chloride are the most preferred, boron trichloride being more preferably.

[0074] In one or more embodiments, selective dealkylation occurs in the presence of a nucleophilic source, such as halogenated hydrochlorides, alkali metal halides, or boron halides. Nucleophilic sources used in selective dealkylation are preferably alkali metal halides, most preferably lithium chloride, sodium bromide, sodium iodide, and potassium iodide, with sodium iodide being the most preferred.

[0075] In one or more embodiments, selective dealkylation occurs in the presence of a polar aprotic solvent, such as dichloromethane (DCM), acetonitrile (ACN), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), and tetrahydrofuran (THF), with acetonitrile and dichloromethane being the most preferred, more preferably dichloromethane.

[0076] In one or more embodiments, selective dealkylation occurs in the concomitant presence of the Lewis acid, the nucleophile source, and the polar aprotic solvent listed here.

[0077] In one or more embodiments, selective dealkylation occurs in the concomitant presence of the Lewis acid and the polar aprotic solvent, without the need for a nucleophilic source. Particularly, when using boron trichloride, it will perform the dual function of Lewis acid and nucleophilic source, making the addition of another nucleophilic source unnecessary.

[0078] The speed of the selective dealkylation reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the process be conducted over a period of 0.5 to 24 hours. In one or more embodiments, the selective dealkylation reaction is carried out at a temperature of 0 °C to 90 °C, preferably 0 °C to 82 °C. In one or more embodiments, the selective dealkylation is carried out at room temperature (20 to 30 °C).

[0079] The selective dealkylation step is particularly advantageous because it enables the reduction of catalytic feedstocks to be used in the asymmetric hydrogenation step. The presence of the hydroxyl group at the 2' position of the substrate of formula III facilitates the reduction, allowing the use of catalytic feedstocks in catalytic hydrogenation equal to or less than 2 mol%, more preferably equal to or less than 1 mol%, and even more preferably less than 1 mol%.

[0080] Regarding the selective dealkylation step, it was observed that the interconversion of rotamers and the proximity of the alkoxy group to be dealkylated to the carbonyl group facilitates chelation by the Lewis acid. The dynamic interconversion between conformations allows the molecule to rapidly explore all possible spatial orientations. This dynamic ensures that the ideal reactive conformation, where the carbonyl oxygen (C=O) and the oxygen of the alkoxy group in the ortho position are in the correct proximity, is readily available to the Lewis acid. In this sense, structures II, Ia, III, and IV can be interchangeably represented by their conformational isomers (rotamers) II', Ia', III', and IV' shown below:

[0081] The alkyloxy substituted isoflavone of formula (IV) can be obtained from the coupling of the substituted 4H-chromen-4-one of formula (V): in which: Each R1, R2, R3, and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy; and X is a halogen; with the substituted phenyl of formula (VI): wherein each R5, R6, R7 and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy; R9 is selected from C1-C1 alkyl; and R10 is a B(OH)2 leaving group. X is preferably selected from fluorine, chlorine, bromine, or iodine, most preferably iodine.

[0082] The term "coupling" refers to the reaction in which two molecular fragments are joined by the formation of a new carbon-carbon covalent bond. The coupling reaction is carried out with the leaving group R10 of boronic acid B(OH)2, and the coupling reaction is called Suzuki coupling.

[0083] All embodiments of R1, R2, R3 and R4 previously performed for compounds (II) and (III) also apply to compound V, including The preferred embodiments of Tables 1 and 2. For compound V, Table 1 can be supplemented by X being a halogen, combining with all the listed embodiments. More preferably, X is iodine, and the preferred compounds V include the radical combinations of Table 2 and X being iodine, and are designated V.1 to V.50, respectively, following the same numbering proposed for compounds II and III. In one or more embodiments, R1, R2, R3, R4, and X are selected such that compound V is V.1, V.2, V.3, V.4, V.5, V.7, V.12, V.17, V.22, V.27, V.32, V.37, V.42, and / or V.47.

[0084] All embodiments of R5, R6, R7, R8, and R9 previously made for compounds (II), (III), and (IV) also apply to compound VI, including the preferred embodiments in Tables 1 and 2. For compound VI, Table 1 can be supplemented by having R9 as a C1-C6 alkyl and R10 as B(OH)2, combining all listed embodiments. More preferably, R9 is methyl and R10 is B(OH)2, and the preferred compounds VI include the radical combinations in Table 2, with R9 being methyl and R10 being B(OH)2, and are designated VI.1 to VI.50, respectively, following the same numbering proposed for compounds II, III, and IV. In one or more embodiments, R5, R6, R7, R8, R9, and R10 are selected such that compound VI is VI.1, VI.2, VI.3, VI.4, VI.5, VI.7, VI.12, VI.17, VI.22, VI.27, VI.32, VI.37, VI.42, and / or VI.47.

[0085] In one or more embodiments, the coupling is carried out in the presence of a palladium catalyst. Preferably, palladium precatalysts are used that are transformed in situ into the active catalytic species. Preferably, the palladium precatalyst is selected from palladium acetate (Pd(OAc)2), tetrakis(triphenylphosphine)palladium(O) (Pd(PPhs)4) or dichlorobis(triphenylphosphine)palladium(II) (PdCl2(PPh3)2), preferably palladium acetate.

[0086] In one or more embodiments, the reaction is carried out under a catalytic loading of 4 mol% or less, preferably 3 mol% or less, and more preferably less than 2 mol%. Preferably, the reaction is carried out without additional phosphine ligands.

[0087] In one or more embodiments, the coupling is carried out in the presence of an inorganic Lewis base, such as carbonates, phosphates, or hydroxides. Inorganic Lewis bases used in the coupling are preferably Carbonates, more preferably alkali metal carbonates, with potassium carbonate being particularly preferred.

[0088] In one or more embodiments, the coupling is carried out in the presence of a polyethylene glycol solvent, such as PEG-200, PEG-300, PEG-400 and PEG-600, with PEG-400 being particularly preferred.

[0089] In one or more embodiments, coupling occurs in the simultaneous presence of the palladium catalyst, the inorganic Lewis base, and the polyethylene glycol solvent listed here.

[0090] The speed of the coupling reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the process be conducted over a period of 1 to 24 hours. In one or more embodiments, the coupling reaction is carried out in a temperature range of 25 °C to 50 °C.

[0091] The substituted 4H-chromen-4-one of formula (V) can be produced from the one-pot two-step reaction of acetophenone of formula (VIII): wherein each R1, R2, R3 and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy; with DMF-DMA for the formation of an enaminone intermediate, and addition of molecular halogen and pyridine for the formation of the substituted 4H-chromen-4-one of formula (V).

[0092] The term “two-step one-pot reaction” refers to a chemical synthesis process where two consecutive reactions occur in the same reaction vessel, without the need to isolate or purify the intermediate between the steps (enaminone). In the terms of the present invention, this definition includes both the process in which all steps occur in a single reactor with Sequential addition of reagents, as well as a continuous series process in which the steps are carried out in interconnected reactors.

[0093] In the first step, acetophenone of formula (VIII) is reacted to form N,N-Dimethylformamide dimethyl acetal (DMF-DMA), being converted into the corresponding enaminone intermediate. In the second step, molecular halogen and pyridine are added to the same reaction medium, allowing the newly formed enaminone to react to form the substituted 4H-chromen-4-one of formula (V).

[0094] All embodiments of R1, R2, R3, and R4 previously made for compounds (II) and (III) also apply to compound VIII, including the preferred embodiments of Tables 1 and 2. More preferably, preferred compounds VIII include the radical combinations of Table 2, and are designated VIII.1 to VIII.50, respectively, following the same numbering proposed for compounds II and III. In one or more embodiments, R1, R2, R3, R4 are selected such that compound V is VIII.1, VIII.2, VIII.3, VIII.4, VIII.5, VIII.7, VIII.12, VIII.17, VIII.22, VIII.27, VIII.32, VIII.37, VIII.42, VIII.47.

[0095] Acetophenone of formula (VIII) can be produced from the acylation of phenol of formula (IX): wherein each R1, R2, R3 and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy; with acetic anhydride in the presence of a Lewis acid.

[0096] The term "acylation" refers to a chemical reaction that introduces an acyl group (RC=O) into a molecule. This reaction forms a new covalent bond between the carbonyl carbon of the acyl group and a nucleophile present in the substrate. in this case the hydroxyl group represented in the phenol of formula (IX). The acylation reaction occurs in the presence of acetic anhydride, thus adding an acetyl radical at the position indicated in compound (VIII). The acylation reaction occurs in the presence of a Lewis acid, such as aluminum trichloride (AlCl3), tin tetrachloride (SnCl4), or borodiethyl ether trifluoride (BF3-OEI2), with borodiethyl ether trifluoride being particularly preferred.

[0097] Particularly for substituted haloalkoxy phenols, they can be derivatized from the reaction of phenols with diethyl bromodifluoromethylphosphonate and sodium hydroxide in an acetonitrile / water mixture. In a preferred embodiment, the reaction starts from a phenolic precursor in an ACN / H2O mixture with a base (KOH). After stirring, the diethyl bromodifluoromethylphosphonate reagent is added, and the reaction proceeds at room temperature for 2 hours.

[0098] All embodiments of R1, R2, R3, and R4 previously made for compounds (II) and (III) also apply to compound IX, including the preferred embodiments of Tables 1 and 2. More preferably, preferred compounds IX include the radical combinations of Table 2, and are designated IX.1 to IX.50, respectively, following the same numbering proposed for compounds II and III. In one or more embodiments, R1, R2, R3, and R4 are selected such that compound IX is IX.1, IX.2, IX.3, IX.4, IX.5, IX.7, IX.12, IX.17, IX.22, IX.27, IX.32, IX.37, IX.42, and / or IX.47.

[0099] In one or more embodiments, the reduced hydroxy-substituted isoflavone of formula II is produced using phenol of formula (IX) as a starting point, which is subjected to acylation to produce acetophenone of formula (VIII). The acetophenone of formula (VIII) is then subjected to a two-step one-pot reaction to form the substituted 4H-chromen-4-one of formula (V), which in turn is coupled by Suzuki coupling to the substituted phenyl of formula (VI) to form the alkyloxy-substituted isoflavone of formula (IV). The alkyloxy-substituted isoflavone of formula (IV) is then subjected to selective dealkylation to form the hydroxy-substituted isoflavone of formula (III), which is then subjected to asymmetric hydrogen transfer for enantioselective formation of the reduced hydroxy-substituted isoflavone of formula (III). (II), by contact with the Noyori-lkariya Ruthenium (S,S) complex in the presence of a hydrogen donor. PROCESS FOR PRODUCING THE (S,S) ISOMER OF A REDUCED HYDROXY-SUBSTITUTED ISOFLAVONE

[0100] In a second aspect, the present invention relates to the production of reduced hydroxy-substituted isoflavone (S,S) of formula (Ha): comprising the asymmetric hydrogen transfer step by contact of hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya complex of Ruthenium (S,S) in the presence of a hydrogen donor, wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy.

[0101] All descriptions and embodiments made in the enantioselective production process of reduced hydroxy-substituted isoflavone also apply to the present aspect of the invention, particularly with regard to the preferred radicals, the Noyori-lkariya complexes of Ruthenium (S,S) and hydrogen donor, as well as all reactions for obtaining compounds (III), (IV), (V), (VI), (VIII) and (IX) and their descriptions.

[0102] All embodiments of R1, R2, R3, R4, R5, R6, R7, and R8 previously made for compounds (II) and (III) also apply to compound Ila, including the preferred embodiments of tables 1 and 2. More preferably, the preferred compounds Ila include the radical combinations of table 2, and are designated lla.1 to Ila.50, respectively, following the same numbering proposed for compounds II and III, as per table 3 below, where OME represents the methoxy radical. Table 3

[0103] In one or more embodiments, R1 , R2, R3, R4, R5, R6, R7 and R8 are selected such that the compound Ha is Ha.1, Ha.2, Ha.3, Ha.4, Ha.5, Ha.7, Ha.12, lla.19, lla.22, lla.27, lla.37, lla. lla.42, and / or lla.47.

[0104] In one or more embodiments, the reduced hydroxy-substituted isoflavone of formula Ha is produced using phenol of formula (IX) as a starting point, which is subjected to acylation to produce acetophenone of formula (VIII). The acetophenone of formula (VIII) is then subjected to a two-step one-pot reaction to form the substituted 4H-chromen-4-one of formula (V), which in turn is coupled, preferably by Suzuki coupling, to the substituted phenyl of formula (VI) to form the alkyloxy-substituted isoflavone of formula (IV). The alkyloxy-substituted isoflavone of formula (IV) is then subjected to selective dealkylation to form the hydroxy-substituted isoflavone of formula (III), which is then subjected to asymmetric hydrogen transfer to form the (S,S) isomer of the reduced hydroxy-substituted isoflavone of formula (Ha), by contact with the Noyori-lkariya ruthenium complex (S,S) in the presence of a hydrogen donor. Enantioselective production process of Pterocarpane - Cyclization in acid medium

[0105] In a third aspect, the present invention relates to a process for producing pterocarpane of formula (I): including the acid cyclization step of the reduced hydroxy substituted isoflavone of formula (II) described herein: wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

[0106] The reduced hydroxy substituted isoflavone of formula (II) is preferably obtained from the enantioselective production process of reduced hydroxy substituted isoflavone described herein. In this way, the reduced hydroxy substituted isoflavone reagent of formula (II) is mainly composed of the (S,S) isomer of formula (Ha), and the pterocarpan obtained is, consequently, mainly composed of the (S,S) isomer of formula 1a.

[0107] In one or more embodiments, after obtaining the reduced hydroxy substituted isoflavone of formula (II) by the enantioselective process described herein, the reduced hydroxy substituted isoflavone (Ha) is isolated and purified, which is then used in the production process of the pretocarpane compound.

[0108] In one or more embodiments, the reduced hydroxy substituted isoflavone of formula (II) used as a reagent has an enantiomeric excess of the (S,S) isomer of at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99% on a molar basis. In one or more embodiments, the resulting pterocarpane compound of formula (I) has an enantiomeric excess of the (S,S) isomer of at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% on a molar basis.

[0109] The term "acid cyclization" refers to a chemical reaction that promotes the formation of a cyclic structure in a molecule, using an acid catalyst. In the present invention, the acid catalyst protonates the hydroxyl group, transforming it into a better leaving group to be attacked by an intramolecular nucleophile, closing the ring and forming a cyclic ether by dehydration.

[0110] All embodiments of R1, R2, R3, R4, R5, R6, R7, and R8 previously made for compounds (II) and (III) also apply to compound I, including the preferred embodiments of tables 1 and 2. More preferably, the preferred compounds I include the radical combinations of table 2, and are designated 1.1 to I.50, respectively, following the same numbering proposed for compounds II and III, as per table 4 below, where OME represents the methoxy radical. Table 4

[0111] In one or more embodiments, R1, R2, R3, R4, R5, R6, R7, and R8 are selected such that compound I is 1.1, I.2, I.3, I.4, I.5, I.7, I.12, I.17, I.22, I.27, I.32, I.37, I.42, and / or I.47.

[0112] In one or more embodiments, the acid cyclization is carried out in the presence of a strong practical acid, preferably hydrochloric, sulfuric, hydrogen bromide or p-toluenesulfonic acid, more preferably hydrochloric acid. In one or more embodiments, the acid cyclization is carried out in the presence of an aprotic solvent such as THF or dioxane, or in the presence of an aliphatic alcohol and ester solvent system, preferably ethanol with ethyl acetate.

[0113] In one or more embodiments, cyclization in an acidic medium occurs in the simultaneous presence of a strong practical acid and a solvent system.

[0114] The speed of the cyclization reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the process be conducted over a period of 5 minutes to 1 hour. In one or more embodiments, the coupling reaction is carried out in a temperature range of 10 °C to 60 °C, preferably at room temperature (20 to 30 °C).

[0115] In one or more embodiments, pterocarpane of formula (I) is produced using phenol of formula (IX) as a starting point, which is subjected to acylation to produce acetophenone of formula (VIII). Acetophenone of formula (VIII) is then subjected to a two-step one-pot reaction to form substituted 4H-chromen-4-one of formula (V), which in turn is coupled by Suzuki coupling to substituted phenyl of formula (VI) to form the substituted alkyloxy isoflavone of formula (IV). The substituted alkyloxy isoflavone of formula (IV) is then subjected to selective dealkylation to form the substituted hydroxy isoflavone of formula (III), which is then subjected to asymmetric hydrogen transfer to enantioselectively form the reduced hydroxy substituted isoflavone of formula (II), by contact with the Noyori-lkariya ruthenium complex (S,S) in the presence of a hydrogen donor. The reduced hydroxy substituted isoflavone of formula (II) It is then subjected to cyclization in an acidic medium to produce the pterocarpane compound of formula (I). PROCESS FOR PRODUCING THE (S,S) ISOMER OF THE PTEROCARPANE COMPOUND - CYCLIZATION IN ACID MEDIUM

[0116] In a fourth aspect, the present invention relates to a process for producing pterocarpane of formula (la): understanding the cyclization step in acid medium of the reduced hydroxy-substituted isoflavone with formula (Ha): where each R1, R2, independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy.

[0117] The reduced hydroxy-substituted isoflavone of formula (Ha) is preferably obtained from the production process of the (S,S) isomer of reduced hydroxy-substituted isoflavone described herein. In this way, the pterocarpane obtained is the (S,S) isomer of formula la.

[0118] In one or more embodiments, after obtaining the (S,S) isomer of the hydroxy-substituted reduced isoflavone of formula (Ha) by the process described herein, the hydroxy-substituted reduced isoflavone is isolated and purified. substituted (Ha), which is then used in the production process of the (S,S) isomer of the pretocarpane compound with formula (la).

[0119] All embodiments of R1, R2, R3, R4, R5, R6, R7, and R8 previously performed for compounds (I), (II), (III), and (Ha) also apply to compound (la), including the preferred embodiments of tables 1, 2, 3, and 4. More preferably, the preferred la compounds include the radical combinations of tables 2, 3, and 4 and are designated la.1 to la.50, respectively, following the same numbering proposed for compounds I, II, III, and Ha, as per table 5 below, where OME represents the methoxy radical. Table 5

[0120] In one or more embodiments, R1, R2, R3, R4, R5, R6, R7 and R8 are selected such that the compound la is la.1, la.2, la.3, la.4, la.5, la.7, la.12, la.17, la.22, la.27, la.32, la.37, la.42 and / or la.47.

[0121] In one or more embodiments, the acid cyclization is carried out in the presence of a strong practical acid, preferably hydrochloric, sulfuric, hydrogen bromide or p-toluenesulfonic acid, more preferably hydrochloric acid. In one or more embodiments, the acid cyclization is carried out in the presence of an aprotic solvent such as THF or dioxane, or in the presence of an aliphatic alcohol and ester solvent system, preferably ethanol with ethyl acetate.

[0122] In one or more embodiments, cyclization in an acidic medium occurs in the simultaneous presence of a strong practical acid and a solvent system.

[0123] The speed of the cyclization reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the process be... conducted over a period of 5 minutes to 1 hour. In one or more embodiments, the coupling reaction is carried out in a temperature range of 10 °C to 60 °C, preferably at room temperature (20 to 30 °C).

[0124] In one or more embodiments, the (S,S) pterocarpane isomer of formula (1a) is produced using phenol of formula (IX) as a starting point, which is subjected to acylation to produce acetophenone of formula (VIII). Acetophenone of formula (VIII) is then subjected to a two-step one-pot reaction to form substituted 4H-chromen-4-one of formula (V), which in turn is coupled by Suzuki coupling to substituted phenyl of formula (VI) to form the substituted alkyloxy isoflavone of formula (IV). The substituted alkyloxy isoflavone of formula (IV) is then subjected to selective dealkylation to form the substituted hydroxy isoflavone of formula (III), which is then subjected to asymmetric hydrogen transfer to form the (S,S) isomer of the reduced hydroxy substituted isoflavone of formula (Ha), by contact with the Noyori-lkariya ruthenium complex (S,S) in the presence of a hydrogen donor.The (S,S) isomer of the reduced hydroxy-substituted isoflavone with formula (Ha) is then subjected to cyclization in an acidic medium to produce the (S,S) isomer of the pterocarpane compound with formula (la). Enantioselective production process of Pterocarpane - Reductive cycling

[0125] In a further aspect, the present invention relates to a process for producing pterocarpane of formula (I): understanding the reductive cyclization step of the hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya ruthenium (S,S) complex in the presence of a hydrogen donor, in which the process is enantioselective for pterocarpane (S,S) of formula (1a): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

[0126] The term "reductive cyclization" refers to a chemical reaction in which a precursor molecule containing at least one pair of functional groups arranged to allow the formation of a ring is subjected to reaction conditions that simultaneously promote an overall reduction of at least one of the functional groups involved or adjacent to the ring and the formation of the new cyclic bond.

[0127] The reductive cyclization of the present invention is an enantioselective process for the (S,S) isomer by utilizing the Noyori-lkariya Ruthenium (S,S) complex, which directs the reaction to preferentially produce the (S,S) enantiomer instead of a racemic mixture. In one or more In these embodiments, the enantiomeric excess of the (S,S) isomer is at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 99%.

[0128] All embodiments of R1, R2, R3, R4, R5, R6, R7, and R8 previously realized for compounds (I), (1a), and (III) also apply to this aspect of the invention, including the preferred embodiments of Tables 1, 2, 4, and 5. More preferably, the preferred compounds I, 1a, and III include the radical combinations of Tables 2, 4, and 5 and are designated 1.1 to I.50, 1a.1 to 1a.50, and 111.1 to III.50, respectively, following the same numbering proposed for compounds I, 1a, and III.

[0129] In one or more concretized forms, R1, R2, R3, R4, R5, R6, R7 and R8 are selected in the form that the compound I is 1.1, I.2, I.3, I.4, I.5, I.7, 1.12, 1.17, I.22, I.27, I.32, I.37, I.42, and / or I.47. In one or more concretizations, R1, R2, R3, R4, R5, R6, R7 and R8 are selected in the form that consists of the following A.1, A.2, A.3, A.4, A.5, A.7, A.12, A.17, A.22, A.27, A.32, A.37, A.42, and / or la.47. In one or more concretized forms, R1, R2, R3, R4, R5, R6, R7 and R8 are selected in the form that consists of 111.1, III.2, III.3, III.4, III.5, III.7, 111.12, 111.17, III.22, III.27, III.32, III.37, III.42, and / or III.47

[0130] The reductive cyclization step occurs in the presence of a hydrogen donor. In one or more embodiments, the hydrogen donor is selected from carboxylic acid salts, carboxylic acids, amines, or a combination thereof. Preferred carboxylic acid salts according to the present invention are C1-C6 carboxylic acid salts. Preferably, the carboxylic acid salts are selected from alkali metal C1-C6 carboxylic acid salts, preferably alkali metal formates. The alkali metals are preferably selected from lithium, sodium, or potassium. In one or more embodiments, the alkali metal C1-C6 carboxylic acid salts are lithium formate, sodium formate, or potassium formate, preferably sodium formate. Carboxylic acids may preferably be selected from C1-C6 carboxylic acids, preferably formic acid.Amines can be selected from C1-C1 trialkylamines, where the term "C1-C1 trialkylamines" refers to tertiary amines that... They possess three alkyl groups, which may be linear or branched chains, and which contain from one to six carbon atoms in total among the three groups. The alkyl groups may be the same or different. Preferred amines include trimethylamine and triethylamine. In one or more embodiments, carboxylic acids and amines are used in combination, in which the amine acts as a co-donor together with the carboxylic acid. Preferably, the combination employed is triethylamine with formic acid.

[0131] The reductive cyclization step occurs in the presence of a Noyori-Ikariya ruthenium (S,S) complex. In one or more embodiments, the Noyori-Ikariya ruthenium (S,S) complex is a Noyori-Ikariya ruthenium (S,S) complex of formula (VII): where M is ruthenium; q6-Ar is an arene ligand of hapticity 6 linked to M selected from optionally substituted arene with Ci-Ce alkyl or Ci-Ce alkoxy; Ph is phenyl; X is halogen; and R13 is arylsulfonyl.

[0132] In one or more embodiments, the optionally substituted arene is selected from the C1-C1e alkyl substituted arene, which may be substituted at one, two, three, four, five, or six carbons of the aromatic ring. Preferably, the C1-C1e alkyl substituted arene is selected from 1-isopropyl-4-methylbenzene (p-cymene), 1,3,5-trimethylbenzene (mesitylene), hexamethylbenzene, and methylbenzene. In one or more embodiments, the arene is unsubstituted (benzene). In one or more embodiments, the halogen is selected from fluorine, chlorine, bromine, or iodine, preferably chlorine. In one or more embodiments, the arylsulfonyl is selected from phenylsulfonyl (Fs), p-toluenesulfonyl (Ts), and naphthalenesulfonyl (Ns).

[0133] The rate of the reductive cyclization reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the process be conducted over a period of 1 hour to 48 hours. In one or more embodiments, the reductive cyclization reaction is carried out in a temperature range of 0 °C to 60 °C.

[0134] In one or more embodiments, the hydroxy-substituted isoflavone of formula (III) is obtained by the steps already described herein, particularly by the selective dealkylation described herein of the alkyloxy-substituted isoflavone of formula (IV). All descriptions and embodiments already made for the selective dealkylation of the alkyloxy-substituted isoflavone of formula (IV) also apply in the present aspect of the invention.

[0135] In one or more embodiments, the alkyloxy substituted isoflavone of formula (IV) is produced from the two-step one-pot reaction of the phenol of formula (IX): wherein each R1, R2, R3 and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy; with the arylacetic acid of formula (X): wherein each R5, R6, R7 and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy, and R9 is selected from C1-C1 alkyl, using a Lewis acid for acylation with the formation of a ketone intermediate in a first step and subsequent formylation / cyclization / dehydration with a polar aprotic solvent and dehydrating agent in a second step.

[0136] The term "one-pot two-step reaction" refers to a chemical synthesis process where two consecutive reactions occur in the same reaction vessel, without the need to isolate or purify the intermediate between the steps (ketone). In the terms of the present invention, this definition includes both the process in which all steps occur in a single reactor with sequential addition of reagents, and also a continuous series process in which the steps are carried out in interconnected reactors.

[0137] All embodiments of R1, R2, R3, and R4 previously made for compounds (II) and (III) also apply to compound IX, including the preferred embodiments of Tables 1 and 2. More preferably, preferred compounds IX include the radical combinations of Table 2, and are designated IX.1 to IX.50, respectively, following the same numbering proposed for compounds II and III. In one or more embodiments, R1, R2, R3, and R4 are selected such that compound IX is IX.1, IX.2, IX.3, IX.4, IX.5, IX.7, IX.12, IX.17, IX.22, IX.27, IX.32, IX.37, IX.42, and / or IX.47.

[0138] All embodiments of R5, R6, R7, R8, and R9 previously made for compounds (II), (III), and (IV) also apply to compound X, including the preferred embodiments in Tables 1 and 2. For compound X, Table 1 can be supplemented by R9 being a C1-C6 alkyl, combining all the listed embodiments. More preferably, R9 is methyl, and the preferred compounds X include the radical combinations in Table 2, R9 being methyl, and are designated X.1 to X.50, respectively, following the same numbering proposed for compounds II, III, and IV. In one or more embodiments, R5, R6, R7, R8, and R9 are selected such that the composite X is X.1, X.2, X.3, X.4, X.5, X.7, X.12, X.17, X.22, X.27, X.32, X.37, X.42, and / or

[0139] In one or more embodiments, the Lewis acid for acylation is selected from metal chloride or boro-diethyl ether trifluoride, preferably boro-diethyl ether trifluoride. Metal chlorides include aluminum chloride, iron(III) chloride, tin chloride, and zinc chloride.

[0140] In one or more embodiments, the polar aprotic solvent is selected from such as dichloromethane, acetonitrile, dimethylformamide, dimethyl sulfoxide and tetrahydrofuran, preferably dimethylformamide.

[0141] In the terms of the present invention, the term “dehydrating agent” refers to any compound, substance or chemical system that is capable of removing a water molecule or equivalent (H2O or HX) from the reaction medium, promoting the formation of a new CO, CN or CC bond through a condensation or elimination reaction, essential for the cyclization and formation of the compound of formula (IV). In one or more embodiments the dehydrating agent is selected from p-toluenesulfonyl chloride or phosphorus oxychloride, preferably p-toluenesulfonyl chloride.

[0142] The rate of the acylation reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the acylation be conducted over a period of 1 to 24 hours. In one or more embodiments, the acylation reaction is carried out in a temperature range of 0 °C to 80 °C.

[0143] The rate of the formylation / cyclization / dehydration reaction is not a limiting factor for the present invention, and it can be carried out over a wide range of reaction times. In one or more embodiments, it is preferable that the formylation / cyclization / dehydration be conducted over a period of 1 to 48 hours. In one or more embodiments, the acylation reaction is carried out in a temperature range of 20 °C to 100 °C.

[0144] In one or more embodiments, pterocarpane of formula (I) is produced using phenol of formula (IX) and acetic acid of formula (X) as starting points, which are subjected to a one-pot two-step acylation and formylation / cyclization / dehydration reaction to produce the substituted alkyloxy isoflavone of formula (IV). The substituted alkyloxy isoflavone of formula (IV) is then subjected to selective dealkylation to form the hydroxy isoflavone. substituted formula (III), which is then subjected to enantioselective reductive cyclization to produce the pterocarpane compound of formula (I), with stoichiometric excess of the (S,S) isomer of formula (1a). PROCESS FOR PRODUCING THE (S,S) ISOMER OF PTEROCARPANE - REDUCTIVE CYCLIZATION

[0145] In a further aspect, the present invention relates to a process for producing pterocarpane of formula (la): understanding the reductive cyclization step of the hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya complex of Ruthenium (S,S) in the presence of a hydrogen donor, wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy.

[0146] All descriptions and embodiments made in the enantioselective production process of pterocarpane of formula (I) by reductive cyclization also apply to the present aspect of the invention, particularly with regard to the preferred radicals, the Noyori-lkariya Ruthenium complexes. (S,S) and hydrogen donor, as well as all the reactions for obtaining compounds (III) and (IV) and their descriptions, in addition to compounds (IX) and (X) and their preferred embodiments.

[0147] All embodiments of R1, R2, R3, R4, R5, R6, R7, and R8 previously carried out for compounds (1a), (III), (IV), (IX), and (X) also apply to the present aspect of the invention, including the preferred embodiments of Tables 1, 2, 3, and 5.

[0148] In one or more embodiments, pterocarpan of formula (1a) is produced using phenol of formula (IX) and acetic acid of formula (X) as starting points, which are subjected to a one-pot two-step acylation and formylation / cyclization / dehydration reaction to produce the substituted alkyloxy isoflavone of formula (IV). The substituted alkyloxy isoflavone of formula (IV) is then subjected to selective dealkylation to form the substituted hydroxy isoflavone of formula (III), which is then subjected to enantioselective reductive cyclization to produce the (S,S) isomer of the pterocarpan compound of formula (1a). Pterocarpane compounds, reduced hydroxy-substituted isoflavones and other intermediates

[0149] In a further aspect, the present invention relates to the pterocarpan compounds of formula (I) and (1a) described herein, as well as the reaction intermediates, including the hydroxy substituted reduced isoflavone of formula (II), the (S,S) isomer of the hydroxy substituted reduced isoflavone of formula (Ha), the 2' hydroxy substituted isoflavone of formula (III), the alkyloxy substituted isoflavone of formula (IV), the 4H-chromen-4-one substituted of formula (V), the phenyl substituted of formula (VI), the acetophenone of formula (VIII), the phenol of formula (IX) and the arylacetic acid of formula (X).

[0150] Preferred compounds according to the present invention include pterocarpan compounds of formula (I) and their (S,S) isomers of formula (1a), the reduced hydroxy-substituted isoflavone of formula (II), the (S,S) isomer of the reduced hydroxy-substituted isoflavone of formula (Ha), the 2'-hydroxy-substituted isoflavone of formula (III) and the alkyloxy-substituted isoflavone of formula (IV).

[0151] All descriptions and embodiments made for such compounds during the detailed description of the synthesis processes of the present invention, particularly with regard to radicals R1, R2, R3, R4, R5, R6, R7, R8, R9, R10 and X, apply to the compounds themselves. Preferred embodiments of the compounds according to the present invention include the embodiments presented in Tables 1, 2, 3, 4 and 5, as well as the derivations of compounds IV, V, VI, VIII, IX and X made from such tables.

[0152] In one or more embodiments, the pterocarpane compound of formula (I) or its S,S isomer of formula (1a) does not include compounds in which R1 is hydrogen, R2 is methoxy, R3 is methoxy, R4 is hydrogen, R5 is hydrogen, R6 is hydrogen, R7 is methoxy, and R8 is hydrogen, simultaneously. Liposomes containing Pterocarpane compound

[0153] In a further aspect, the present invention relates to a liposome comprising lipids and a pterocarpan compound. Pterocarpans are known in the art and possess a tricyclic skeleton where the central aromatic ring is fused to a dihydrofuran ring via a COC linkage, giving it the benzofuran-chroman structure. In one or more embodiments, the liposome comprises from 2% to 10% by weight of pterocarpan compound, based on the total weight of the liposome.

[0154] In one or more embodiments, the pterocarpane compound is the pterocarpane compound described herein, having formula (I): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

[0155] Particularly with regard to compound (I), all descriptions of R1, R2, R3, R4, R5, R6, R7, and R8 previously given for compound (I) also apply to the liposome of the present invention, including the preferred embodiments in Tables 1 and 4. More preferably, the preferred compounds I include the radical combinations in Table 4 designated 1.1 to 1.50.

[0156] In one or more embodiments, the pterocarpan compound of formula (I) is in its dextrorotatory (+), levorotatory (-) form or in a stereoisomeric or racemic mixture thereof. In one or more embodiments, the pterocarpan compound of formula (I) is in a stereoisomeric mixture enriched with its dextrorotatory (+) form of formula (1a), the S,S isomer. In one or more embodiments, the pterocarpan compound of formula (I) is in its dextrorotatory (+) form of formula (1a), the S,S isomer.

[0157] Particularly with regard to compound (la), all descriptions of R1, R2, R3, R4, R5, R6, R7, and R8 previously made for compounds (la) also apply to the liposome of the present invention, including the preferred embodiments of tables 1 and 5. More preferably, the preferred compounds la include the radical combinations of table 5 designated la.1 to la.50.

[0158] The term "lipids" refers to the amphiphilic and structural compounds that form the vesicular bilayer, encompassing not only the primary phospholipids that constitute the fundamental matrix of the membrane, but also accessory, rigidity, and stabilizing agents. In one or more embodiments, lipids are selected from phospholipids, cholesterol, or a combination thereof. Phospholipids include primary phospholipids, which constitute the major structural part of the lipid bilayer, and accessory phospholipids for modification or optimization of physicochemical properties. Phospholipids can be selected from phosphatidylcholines, phosphatidylglycerols, phosphatidylserine, phosphatidylinositol, distearolphosphatidylethanolamine, dipalmitoylphosphatidylcholine, dileoylphosphatidylethanolamine, or a combination thereof. Phosphatidylcholines are preferred as primary structural phospholipids.Phosphatidylglycerols can be used either as primary phospholipids or as a phospholipid. Phosphatidylserine and phosphatidylinositol can be used as accessory phospholipids, responsible for adding a negative surface charge or mimicking properties of biological membranes for cell recognition. Distearoylphosphatidylethanolamine (DSPE) serves as a base for the conjugation of stealth agents, promoting the formation of stealth liposomes. Dipalmitoylphosphatidylcholine (DPPC) has a phase transition temperature around 41 °C, making it compatible with the development of thermosensitive liposomes. Dioleoylphosphatidylethanolamine (DOPE) is used for the development of pH-sensitive liposomes. In one or more embodiments, the liposome comprises phosphatidylcholine as the primary phospholipid and phosphatidylserine as the accessory phospholipid. In one or more embodiments, the liposome of the present invention comprises from 45 mol% to 80 mol% of phospholipids, based on total moles of lipids.

[0159] In one or more embodiments, the liposome comprises cholesterol. Its inclusion in the liposome formulation is capable of modulating membrane fluidity, increasing liposome rigidity and stability and decreasing vesicle permeability, optimizing the retention of the encapsulated pterocarpan. In one or more embodiments, the liposome of the present invention comprises from 10 mol% to 45 mol% cholesterol, based on total lipid moles.

[0160] In one or more embodiments, the liposome according to the present invention further comprises a stealth agent. The term "stealth agent" refers to components added to the surface of the nanoparticle with the primary purpose of avoiding recognition and removal by the body's immune system. Useful stealth agents according to the present invention may be selected from polyethylene glycol, polysarcin, polycarboxybetaine, or a combination thereof, preferably polyethylene glycol. The polyethylene glycol according to the present invention may have varying average molecular weights, preferably in the range of 1000 to 5000 Da, more preferably 2000 (PEG-2000). In one or more embodiments, the liposome of the present invention comprises from 1 mol% to 5 mol% by weight of stealth agent, based on the total moles of lipids.

[0161] In one or more embodiments, the liposome further comprises a conjugating agent. The term conjugating agent refers to a molecule used to establish a stable covalent bond between the lipid bilayer of the liposome and a desired external functional group. In one or more embodiments, the conjugating agent is selected from maleimide, iodoacetamide, n-hydroxysuccinimide, and carbodiimide, or a combination thereof, preferably maleimide. In one or more embodiments, the liposome of the present invention comprises from 1 mol% to 5 mol% by weight of conjugating agent, based on the total moles of lipids.

[0162] The stealth agent and the conjugating agent may be present in the liposome concomitantly in the form of a bifunctional conjugating agent, preferably polyethylene glycol-maleimide (PEG-MAL), more preferably PEG2000-MAL. In one or more embodiments, the stealth agent and the conjugating agent are present in the liposome concomitantly in the form of a heterobifunctional conjugating agent, comprising an anchoring agent, the stealth agent, and the conjugating agent. The term "anchoring agent" refers to any component with a substantial lipophilic nature and compatible with the liposome bilayer, whose main function is to spontaneously insert itself into the liposome structure. This agent stabilizes the stealth agent and the conjugating agent that extend outward from the liposome surface.Useful anchoring agents for the present invention include the phospholipids and cholesterol listed herein, preferably distearoyl phosphatidylethanolamine (DSPE) and dipalmitoyl phosphatidylethanolamine, with distearoyl phosphatidylethanolamine being more preferably the latter. In one or more embodiments, the anchoring agent and the conjugating agent are present in the liposome concomitantly in the form of a heterobifunctional DSPE-PEG-MAL conjugating agent, preferably DSPE-PEG2000-MAL. In one or more embodiments, the liposome of the present invention comprises from 1% to 5% by weight of bifunctional or heterobifunctional agent, based on total moles of lipids.

[0163] In one or more embodiments, the liposome has a drug-to-lipid ratio at a lower limit of at least 2%, at least 2.5%, at least 4%, preferably at least 5%, and an upper limit of up to 10%, up to 12%, up to 15%, up to 20%, where each lower limit can be combined with each upper limit to define ranges.

[0164] The term "drug-lipid ratio" (D / L) refers to the mass ratio of the active ingredient (drug), particularly pterocarpan compound, to the total mass of the lipid components of the liposome. This ratio is usually expressed as a mass ratio (drug mass:total lipid mass), or as a mass percentage, according to the equation below: drug mass - - - - , — X 100% drug mass + total lipid mass

[0165] The drug-to-lipid ratio is particularly relevant because it directly influences encapsulation efficiency (the liposome's ability to retain the drug), the physicochemical stability of the liposomal vesicle, and consequently, the dose of drug that can be safely and effectively administered. An ideal D / L ratio maximizes drug loading without compromising the structural integrity of the liposome, ensuring a therapeutically relevant and economically viable end product.

[0166] In the context of the present invention, the term "total lipid mass" refers not only to the mass of the liposome lipids, but also to the mass of any stealth, conjugating and anchoring agents, or other components other than the active compound pterocarpan, added to the liposome formulation.

[0167] Similarly, the term “total moles of lipids” as used herein refers to the total moles not only of the liposome lipids, but also to the moles of any stealth, conjugating and anchoring agents, or other components besides the active compound pterocarpan, added to the liposome formulation. The denotation of liposome constituents on a molar basis is particularly used in the art.

[0168] The term “total liposome weight” as used herein refers to the total weight of the liposome lipids, any stealth, conjugating and anchoring agents, or other components, and also the active compound pterocarpan, added to the liposome formulation.

[0169] In one or more embodiments, the liposome of the present invention comprises phosphatidylcholine and phosphatidylserine as phospholipids, in addition to cholesterol, DSPE-PEG2000-MAL and a pterocarpane compound.

[0170] In one or more embodiments, the liposome is an immunoliposome functionalized with a monoclonal antibody. The term immunoliposome refers to a liposomal system that has been modified or conjugated on its outer surface with a biological recognition molecule, particularly an antibody. The monoclonal antibody referred to here is a biological recognition protein obtained by genetic engineering techniques. For the purposes of conjugation to the liposomal nanosystem, antibodies should not be limited to a specific type or class, provided they contain free thiol groups, such as those found in cysteine ​​residues, or via a thiolization process that transforms primary amines present in the antibody's lysine residues into thiol groups (-SH), to facilitate covalent anchoring to the conjugating agent.In one or more embodiments, the monoclonal antibody is an anti-epidermal growth factor receptor (anti-EGFR) agent, with cetuximab being a non-limiting example of this preferred class of antibodies.

[0171] In one or more embodiments, the liposome or immunoliposome has a size in the range of 80 to 150 nm, measured by dynamic light scattering (DLS). It is preferable that the liposome or immunoliposome has a zeta potential as far away from zero mV as possible, with values ​​of up to -15 mV being preferable, measured by electrophoretic light scattering (ELS). In one or more embodiments, the liposome or immunoliposome has a polydispersity index (PDI) of less than 0.25, measured by dynamic light scattering (DLS). Liposome Production Process In a further aspect, the present invention relates to a liposome production process described herein by means of the lipid film hydration method. In one or more embodiments, the liposome production process comprises the steps of: a) dilution of the total lipid mass and pterocarpan compound in a volatile organic solvent; b) formation of a lipid film; c) evaporation of the volatile organic solvent; d) solubilization of the lipid film in aqueous phase; and e) fragmentation to obtain single-layer nanoparticles.

[0172] The dilution step of the total lipid mass and pterocarpane compound in a volatile organic solvent comprises the complete dissolution of all liposome components in a volatile organic solvent, particularly one with a boiling point below 80 °C, preferably below 70 °C at atmospheric pressure. Useful solvents for the present invention include, but are not limited to, chloroform, methanol, ethanol, diethyl ether, tetrahydrofuran, cyclohexane, hexane, or a combination thereof, preferably chloroform. The complete miscibility of the components in the organic phase allows the lipid molecules to mix intimately before their organization into the bilayer.

[0173] The term “total lipid mass,” as defined herein, refers not only to the lipids of the liposome, but also to any stealth, conjugating, and anchoring agents, or other components besides the pterocarpan compound, added to the liposome formulation. All descriptions and qualitative and quantitative embodiments of all such components provided in the detailed description of the liposome also apply to the present aspect of the invention.

[0174] In one or more embodiments, the pterocarpan compound and the lipid mass are dissolved in a drug-to-lipid ratio at a lower limit of at least 2%, at least 2.5%, at least 4%, preferably at least 5%, and an upper limit of up to 10%, up to 12%, up to 15%, up to 20%, wherein each lower limit may be combined with each upper limit for range definition.

[0175] The steps of lipid film formation and subsequent evaporation of the organic solvent are interconnected, aiming to remove the volatile organic solvent to force the lipids into an anhydrous and highly organized state. The process consists of spreading the homogeneous lipid solution obtained in step (a) on a surface, followed by the gradual and complete removal of the volatile organic solvent. The result is the formation of a thin and uniform lipid layer on the surface. The dry film serves as the precursor matrix for the self-organization of vesicular structures in the subsequent hydration step. The techniques used for the evaporation step are known in the art and are not particularly limited; examples include rotary evaporators, thin-film evaporators, plate evaporators, and spray drying.

[0176] The aqueous phase lipid film solubilization step (hydration) is a spontaneous self-organization process in which the anhydrous lipid film is exposed to an aqueous solution, usually a neutral pH (6 to 8) aqueous buffer. Aqueous buffers are not particularly limited as long as they control the pH and maintain the correct osmolarity. Useful buffers for the present invention are not limited to, but may be selected from, phosphate buffer (PB), phosphate-buffered saline (PBS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, or Tris(hydroxymethyl)aminomethane (TRIS) buffer, preferably phosphate buffer (PB). The amphiphilic nature of lipids, combined with the free energy of the interface, causes them to thermodynamically self-organize into concentric lamellar structures, forming multilamellar vesicles. Hydration results in structures containing discrete and isolated aqueous compartments.

[0177] The fragmentation step is a process of applying mechanical or ultrasonic energy to the suspension of multilamellar vesicles obtained in step d). This energy promotes the fragmentation of the large multilamellar vesicles, resulting in the formation of smaller, more homogeneous unilamellar nanoparticles. The techniques used for fragmentation are not particularly limited and can be selected from sonication, either by probe tip or ultrasonic bath, as well as methods involving the application of high-energy mechanical or shear forces. Such methods include extrusion, high-pressure homogenization, and solvent injection, such as ethanol injection.

[0178] In one or more embodiments, the liposome production process additionally includes step f) solution purification. The purification step aims to remove unwanted components and concentrate the liposomal nanoparticles. This step involves the elimination of compounds Unencapsulated (free) pterocarpane, residues of the organic solvent used, and any other chemicals introduced during the preparation or fragmentation phases. The techniques used for purifying the liposomal solution are not restricted and should be selected based on the scale of production, stability, and characteristics of the formulation. Examples of purification techniques include, but are not limited to, filtration, dialysis, size exclusion chromatography (SEC), centrifugation, and tangential flow ultrafiltration / diafiltration.

[0179] In one or more embodiments, the liposome production process further comprises step g) of conjugation of the monoclonal antibody to the liposome. Methods of liposome-antibody conjugation are known in the art and are described, for example, in Eloy, JO, Petrilli, R., Chesca, D. L, Saggioro, F. R, Lee, RJ, & Marchetti, JM (2017). Anti-HER2 immunology for co-delivery of paclitaxel and rapamine for breast cancer therapy. European Journal of Pharmacology and Biopharmaceutics, 115, 159-167, incorporated herein by reference. The descriptions and embodiments of monoclonal antibodies carried out in the detailed description of the liposome also apply to the present aspect of the invention. PHARMACEUTICAL COMPOSITION

[0180] In a further aspect, the present invention relates to a pharmaceutical composition comprising pterocarpan compound of formula (I) described herein, or the liposome described herein. All descriptions and embodiments of the pterocarpan compound and the liposome already made also apply to the present aspect of the invention.

[0181] In one or more embodiments, the pharmaceutical composition additionally comprises a pharmaceutically acceptable vehicle, diluent and / or excipient. The term "acceptable" or "pharmaceutically acceptable," with respect to a formulation, composition or ingredient, as used herein, means that it has no persistent detrimental effect on the general health of the individual being treated or does not negate the biological activity or properties of the compound, and is relatively non-toxic.

[0182] Pharmaceutical compositions can be formulated conventionally, using one or more physiologically acceptable carriers. including excipients and auxiliaries, which facilitate the processing of the active compound into preparations that can be used pharmaceutically. The appropriate formulation depends on the chosen route of administration. Any of the well-known techniques, carriers, and excipients may be used as appropriate and understood in the art. A summary of the pharmaceutical compositions described herein may be found, for example, in Remington: The Science and Practice of Pharmacy, Nineteenth Ed. (Easton, PA: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999), which are incorporated herein by reference in their entirety.

[0183] A pharmaceutical composition, as used herein, refers to a mixture of the pterocarpan compound described herein, the liposome or immunoliposome described herein with other chemical components, such as carriers, stabilizers, diluents, dispersing agents, suspending agents, thickening agents and / or excipients. The pharmaceutical composition facilitates the administration of the compound, liposome or immunoliposome to an organism. In the practice of the treatment or use methods provided herein, therapeutically effective amounts of the compound, liposome or immunoliposome described herein are administered in a pharmaceutical composition to an individual with a disease, disorder or condition to be treated. Preferably, the individual is a mammal, and preferably the mammal is a human being. A therapeutically effective amount may vary widely depending on the severity of the disease, the age and relative health status of the individual, the potency of the compound used and other factors.The compound, liposome, or immunoliposome can be used alone or in combination with one or more therapeutic agents as components of mixtures.

[0184] The term "therapeutically effective amount," as used herein, refers to a sufficient quantity of an administered agent, compound, liposome, or immunoliposome that will alleviate, to some extent, one or more... symptoms of the disease being treated. The result may be the reduction and / or relief of the signs, symptoms, or causes of the disease, or any other desired alteration of a biological system. The term "therapeutically effective amount" includes, for example, a prophylactically effective amount. An "effective amount" of a compound, liposome, or immunoliposome disclosed herein is an amount effective to achieve a desired pharmacological effect or therapeutic improvement without undue adverse side effects. It is understood that "an effective amount" or "a therapeutically effective amount" may vary from individual to individual due to variation in the metabolism of the compound / liposome / immunoliposome, age, weight, general condition of the individual, the condition being treated, the severity of the condition being treated, and the judgment of the prescribing physician.By way of example only, therapeutically effective amounts can be determined through routine experimentation, including but not limited to a dose-escalation clinical trial.

[0185] In certain embodiments, the compositions may also include one or more pH adjusting agents or buffering agents, including acids such as acetic, boric, citric, lactic, phosphoric, and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane; and buffers such as citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in an amount necessary to maintain the pH of the composition within an acceptable range.

[0186] In other embodiments, the compositions may also include one or more salts in an amount necessary to bring the osmolality of the composition into an acceptable range. Such salts include those possessing sodium, potassium, or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0187] The pharmaceutical compositions described herein can be administered to an individual via multiple routes of administration, including, but not limited to, oral, parenteral (e.g., intravenous), intraperitoneal, subcutaneous, intramuscular, intranasal, buccal, topical, rectal, or transdermal routes. Preferably intravenous or intraperitoneal. The pharmaceutical formulations described herein include, among others, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal or immunoliposomal dispersions, aerosols, solid dosage forms, powders, immediate-release formulations, controlled-release formulations, rapid-melting formulations, tablets, capsules, dragees, delayed-release formulations, prolonged-release formulations, pulsatile-release formulations, multiparticulate formulations, and mixed immediate-release and controlled-release formulations.

[0188] Pharmaceutical compositions that include a compound described herein may be manufactured in conventional ways, such as, but not limited to, conventional mixing, dissolution, granulation, coating of dragees, levigation, emulsification, encapsulation, trapping or compression. Uses of the compound Pterocarpan, Liposome, and Pharmaceutical Composition

[0189] In a further aspect, the present invention relates to the use of the pterocarpan compound of formula (I) described herein, the liposome described herein or the pharmaceutical composition described herein in the manufacture of a medicament for treating cancer. All descriptions and embodiments of the compound, liposome and pharmaceutical composition already made also apply to the present aspect of the invention.

[0190] The term "cancer" refers to or describes the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, breast cancer, prostate cancer, leukemia, myelodysplastic syndrome, glioma, colon cancer, lung cancer, and ovarian cancer, preferably prostate cancer. In one or more embodiments, cancer is selected from the group consisting of breast cancer, prostate cancer, leukemia, myelodysplastic syndrome, glioma, colon cancer, lung cancer, and ovarian cancer, preferably prostate cancer.

[0191] The terms "treat" or "treatment," as used herein, include relieving, alleviating, or improving a disease or condition, or its symptoms; To control a disease or condition, or its symptoms; to prevent additional symptoms; to attenuate or prevent the underlying metabolic causes of the symptoms; to inhibit the disease or condition, for example, to stop the development of the disease or condition; to alleviate the disease or condition; to cause the regression of the disease or condition; to alleviate a condition caused by the disease or condition; or to stop the symptoms of the disease or condition. The terms "treat" or "treatment" include, but are not limited to, prophylactic and / or therapeutic treatments.

[0192] In one or more embodiments, the medicine is manufactured for administration to an individual by multiple routes of administration, including, but not limited to, oral, parenteral (e.g., intravenous), intraperitoneal, subcutaneous, intramuscular, intranasal, buccal, topical, rectal, or transdermal routes of administration, preferably intravenous or intraperitoneal. TREATMENT METHOD

[0193] In a further aspect, the present invention relates to a treatment method comprising administering the pterocarpan compound of formula (I) described herein, the liposome described herein, or the pharmaceutical composition described herein to an individual with cancer. All descriptions and embodiments of the compound, liposome, and pharmaceutical composition already made also apply to the present aspect of the invention.

[0194] In one or more embodiments, the cancer is selected from the group consisting of breast cancer, prostate cancer, leukemia, myelodysplastic syndrome, glioma, colon cancer, lung cancer, and ovarian cancer, preferably prostate cancer.

[0195] In one or more embodiments, treatment is administered to an individual via multiple routes of administration, including, but not limited to, oral, parenteral (e.g., intravenous), intraperitoneal, subcutaneous, intramuscular, intranasal, buccal, topical, rectal, or transdermal routes, preferably intravenous or intraperitoneal. In one or more embodiments, the drug is administered intravenously or intraperitoneally. EXAMPLES

[0196] The present invention will be better understood by means of the following examples, which are not limiting to the invention described herein. Example 1 - Chemical synthesis and obtaining of pterocarpane compound (Route 1)

[0197] In the first synthesis step of the pterocarpane compound (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane), the acylation of 3,4-dimethoxyphenol with acetic anhydride was carried out using BF3.OEI2 as a Lewis acid, resulting in the corresponding acetophenone (1-(2-hydroxy-4,5-dimethoxypheni)ethan-1-one). In a mixture of 3,4-dimethoxyphenol and acetic anhydride, the catalyst BF3Et2O is added at 0 °C. Subsequently, the reaction is heated to 90 °C and maintained under stirring for 1 hour. A yield of 95% was obtained.

[0198] Next, in the second stage, two one-pot reactions were applied: one being the reaction of acetophenone with DMF-DMA to form an intermediate enaminone (not isolated), and the other being the addition of molecular iodine and pyridine, obtaining 3-iodo-6,7-dimethoxy-4H-chromen-4-one. The formation of the enaminone is carried out by solubilization in toluene with DMF-DMA and the mixture under reflux at 110 °C for 12 hours. The toluene is removed and the product of the first reaction is redissolved in chloroform, then pyridine and iodine (I2) are added and the mixture is kept under stirring at room temperature for 3 hours. A yield of 93% was obtained.

[0199] Next, in the third step, the Suzuki coupling reaction was carried out between 3-iodo-6,7-dimethoxy-4H-chromen-4-one and 2,4-dimethoxyphenylboronic acid (1.5 equivalent), with a 2% molar Pd catalyst (Pd(OAc)2) in the presence of the inorganic base potassium carbonate (K2CO3) and PEG-400 solvent, at 50 °C, forming the isoflavone (3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one), as shown in the scheme below. A yield of 99% was obtained. PEG-400, 4 h

[0200] Then, in the fourth step, selective demethylation of the isoflavone (3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one) was carried out with aluminum chloride, using 3 equivalents of AlCl3 and 2 equivalents of NaI under reflux with acetonitrile (ACN) for 18 hours at room temperature, resulting in 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4 / - / -chromen-4-one, as shown in the scheme below. A yield of 71% was obtained.

[0201] Selective demethylation was also carried out, in a second embodiment, using BCl3 (1M in hexane) in anhydrous dichloromethane at room temperature for 1 hour. In this embodiment, a yield of 90% was achieved. Thus, it was possible to perform selective demethylation in less time and with a higher yield when compared to the first embodiment.

[0202] Next, in the fifth step, the asymmetric hydrogen transfer reaction via dynamic kinetic resolution (ATH-DKR) catalyzed by the Noyori-lkariya ruthenium complex (S,S) was carried out, with the q6-Ar group being p-cymene, the halogen being chlorine, and the arylsulfonyl group being p-toluenesulfonyl, at 2 molar percent. HCO2Na was used as the hydrogen source, CTAB as the surfactant, and methanol as the solvent. The reaction was carried out at 45 °C for 18 hours. Through one-pot reduction of C=C and C=O bonds of 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4 / - / -chromen-4-one, the enantiomerically enriched cis alcohol (S,S), (3S,4S)-3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxychroman-4-ol, is formed, as shown in the scheme below. A yield of 99% was obtained, with an enantiomeric excess of the (S,S) isomer of 98%. 45°C, 6 PM (3S,4S)

[0203] In particular, the synthesis of the present invention shows a surprising reproducibility of the fifth-step asymmetric hydrogen transfer reactions employing 2 mol% catalytic feedstock in the scale-up from 25-30 mg to 250-1000 mg.

[0204] Next, in the sixth step, the pterocarpane compound (+)-(6aS,11aS)-2,3,9-trimethoxy-6a,11a-dihydro-6H-benzofuro[3,2-c]chromene, also called (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane ((+)-PTC), was then obtained by cyclization of (3S,4S)-3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxychroman-4-ol in an acidic medium, using cHCl (6 equiv.) as a strong practical acid, EtOH / AcOEt (1:2) as solvent, at a temperature of 25 °C for 7 minutes, according to the scheme below. A yield of 82% was obtained. (3S.4S)

[0205] From the six steps performed, an overall yield of 50.4% was observed when using selective demethylation with AlCh, NaI, and ACN, and an overall yield of 63.8% when performing selective demethylation with BCl3 and dichloromethane.

[0206] The compounds (1-(2-hydroxy-4,5-dimethoxyphenyl)ethan-1-one), obtained in the first step, 3-iodo-6,7-dimethoxy-4H-chromen-4-one, obtained in the second step, (3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one), obtained in the third step, 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4 / - / -chromen-4-one, obtained in the fourth step, (+)-(6aS,11aS)-2,3,9-trimethoxy-6a,11a-dihydro-6H-benzofuro[3,2-c]chromene, obtained in the fifth step, and (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane, obtained in the sixth step, were analyzed by nuclear magnetic resonance (NMR) of 1H and the resulting spectra are presented in Figures 1, 2, 3, 5, 6, and 7, respectively. The intermediate compound (3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one), obtained in the third step, was also analyzed by NMR of 13 C and the resulting spectrum is shown in Figure 4.

[0207] 0 compounds (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane and (-)-(6aS,11aS)-2,3,9-trimethoxypterocarpane were analyzed by high-performance chromatography and the results are presented in Figures 8 and 9.

[0208] With the synthesis developed here, pterocarpan compounds can be obtained enantioselectively for the (S,S) isomer with significantly higher yields compared to syntheses described in the prior art. The yield ranges reported here are considered high for a 6-step synthesis, highlighting the high yields obtained in the individual steps.

[0209] The robustness of the asymmetric hydrogen transfer methodology and the ease of obtaining isoflavones allows for the production of new derivatives, which contribute to the establishment of new structure-activity relationships, through the production of (+)-PTC derivatives with different substitution patterns, as established in the Markush formulas of the present invention and in the embodiments of Tables 1 to 5. Example 2 - Chemical synthesis and obtaining of pterocarpane compound (Route 2)

[0210] In the first synthesis step of the pterocarpane compound (+)-(6aS,11aS)-2,3,9-trimethoxypterocarpane), a Friedel-Crafts acylation of 3,4-dimethoxyphenol was performed using BF3*Et2O-promoted 2,4-dimethoxyphenyl acetic acid, commercially derived, as the acyl residue source. The intermediate ketone was obtained using this method, and treatment with DMF in the presence of p-TsCl resulted in the predicted formylation / cyclization / dehydration reaction sequence, leading to the release of 3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one with a 73% yield, as shown in the scheme below:

[0211] A magnetically stirred mixture of 3,4-dimethoxyphenol (2.31 g, 15.0 mmol, 1.5 equiv.) and 2,4-dimethoxyphenylacetic acid (1.96 g, 10.0 mmol) in diethyl boron trifluoride etherate (7.4 mL, 60.0 mmol, 6.0 equiv.) contained in a 250 mL round-bottom flask was stirred at 65 °C (oil bath) for 7 h. The resulting mixture was cooled in an ice bath and then treated dropwise with DMF (15.4 ml) before being heated to 22 °C and, after a further 0.25 h, then heated to 50 °C. Next, p-toluenesulfonyl chloride (5.7 g, 30.0 mmol, 3.0 equiv.) was added to the reaction mixture, which was then heated to 75 °C for 17.5 h. The cooled reaction mixture was extracted with EtOAc (1 x 350 ml) and the combined organic phases were then washed with water (2 x 60 ml) and brine (1 x 80 ml) before being dried (Na2SO4), filtered, and concentrated under reduced pressure. The resulting residue was subjected to flash chromatography (silica, petroleum ether / ethyl acetate elution 5:4 v / v) to provide, after concentration of the appropriate fractions (Rf = 0.5), the compound 3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (2.50 g, 73%) as a whitish solid.A small portion of the intermediate fractions from this chromatographic purification process was concentrated under reduced pressure to provide a light brown solid, mp = 181-183 °C. 1H NMR (500 MHz, CDCl3) õ 7.89 (s, 1H), 7.61 (s, 1H), 7.24 (d, J = 9.0 Hz, 1H), 6.86 (s, 1H), 6.56-6.54 (m complex, 2H), 3.97 (s, 3H), 3.96 (s, 3H), 3.83 (s, 3H), 3.77 (s, 3H); 13C{1H} NMR (125 MHz, CDCI3) õ 175.7, 161.2, 158.7, 154.3, 153.6, 152.4, 147.6, 132.3, 121.9, 118.0, 113.8, 105.1, 104.6, 99.7, 99.2, 56.5, 56.4, 55.8, 55.5; IR Umax 3357, 2919, 2850, 1632, 1609, 1509, 1470, 1271, 1207, 1024, 966, 817, 790 cm-1; HRMS (ESI, +ve) m / z [M+H]+ calculated for C19H18O6 343.1176; Found 343.1177.

[0212] In the second synthesis step, the selective demethylation of 3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one was performed by treating this tetramethoxylated isoflavone with a mixture of AIDS and sodium iodide in acetonitrile, obtaining the corresponding trimethoxylated 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one with a yield of 73%. The selectivity of this step derives from the simultaneous complexation of the Lewis acid (AlCl3) with the carbonyl oxygen and the oxygen of the nearby methoxy group within the rotameric form of the substrate, and with the iodide ion then attacking the associated methyl group to provide the observed product, as shown in the scheme below:

[0213] A solution of the compound 3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (50 mg, 0.147 mmol) in ACN (1.0 mL) magnetically stirred was cooled to 0 °C (ice water bath) and then treated with AlCl3 (59 mg, 0.441 mmol, 3 equiv.) and NaI (44 mg, 0.294 mmol, 2 equiv.). The resulting mixture was then heated (oil bath) under reflux for 16 h before being cooled to room temperature, poured into ice water (5 mL) and diluted with EtOAc (25 mL). The separated organic phase was washed with brine (1 x 5 mL) before being dried (Na2SO4), filtered and concentrated under reduced pressure. The resulting residue was subjected to flash chromatography (silica, petroleum ether / ethyl acetate elution 3:2 v / v) to provide, after concentration of the appropriate fractions (Rf = 0.5), the compound 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (36 mg, 73%) as a white crystalline solid, mp = 210-212 °C.RMN de 1 H (500 MHz, CDCI3) õ 9,42 (s, 1 H), 8,07 (s, 1 H), 7,64 (s, 1 H), 7,07 (d, J = 8,5 Hz, 1H), 6,94 (s, 1H), 6,65 (d, J = 2,5 Hz, 2H), 6,54 (dd, J = 8,5 e 2,5 Hz, 1 H), 4,02 (s, 3H), 4,01 (s, 3H), 3,82 (s, 3H); 13C{1 H} RMN (125 MHz, CDCI3) õ 178,4, 162,1, 158,2, 155,5, 154,4, 148,5, 130,4, 124,6, 116,8, 113,2, 107,8, 104,6, 104,5, 99,3, 56,6, 55,5; IR umax 3338, 1624, 1506, 1431, 1274, 1225, 1161, 1026 cm-1; HRMS (ESI, +ve) m / z [M+H]+ calculado para C18H16O6 329,1019; Encontrado 329,1019.

[0214] In the third step, the enantioselective reductive cyclization of 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one was carried out using a combination of formic acid and triethylamine in the presence of the commercially available Noyori-Ikariya catalyst (S,S)Ru-TsDPEN, whose q6-Ar group is p-cymene, the halogen is chlorine, and the arylsulfonyl group is p-toluenesulfonyl, using DMSO as solvent. Under these conditions, and when this reduction was carried out at 45 °C for 24 h, (+)-(6aS,11aS)-2,3,9-trimethoxy-6a,11a-dihydro-6H-benzofuro[3,2-c]chromene was obtained as an oil with a yield of 52% and an enantiomeric excess (ee) of 96%, as determined by chiral HPLC.

[0215] Magnetically stirred triethylamine (900 pL) was cooled to 0 °C under an argon atmosphere, then treated with formic acid (300 pL), and the resulting solution was heated to room temperature and after 5 minutes treated with a solution of RuCl(p-cymene)[(S,S)-Ts-DPEN] (23 mg, 0.036 mmol, 0.036 equivalents) in DMSO (440 pL). After a further 5 min, a solution of 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (328 mg, 1.0 mmol) in DMSO (1.25 ml) was added to the reaction mixture, which was then heated to 45 °C (oil bath) for 24 h. Next, the reaction mixture was cooled to room temperature and then treated with hydrochloric acid (520 µL of 37% aqueous solution). After a further 10 min, the reaction mixture was diluted with NH4Cl (30 mL of a saturated aqueous solution) before being extracted with EtOAc (3 x 30 mL). The combined organic phases were then dried (NaSO4), filtered, and concentrated under reduced pressure.The resulting residue was subjected to flash chromatography (silica, petroleum ether / ethyl acetate elution 5:1 v / v) to give, after concentration of the appropriate fractions (Rf = 0.2), the compound (+)-(6aS, 11aS)-2,3,9-trimethoxy-6a, 11a-dihydro-6H-benzofuro[3,2-c]chromene (163 mg, 52%) as a clear, colorless oil, [a]D = +184° (c = 0.009, EtOH) {lit. [a]D = +203° (c = 1.3, EtOH)}. mp = 110-111 °C (lit. mp = 115-118 °C). 1H NMR (500 MHz, CDCl3) δ 7.12 (d, J = 9.0 Hz, 1H), 6.97 (s, 1H), 6.48 (s, 1H), 6.46- 6.44 (m complex, 2H), 5.48 (d, J = 7.0 Hz, 1 H), 4.22 (dd, J = 10.5 and 4.5 Hz, 1 H), 3.89 (s, 3H), 3.84 (s, 3H), 3.76 (s, 3H), 3.58 (t, J = 11.0 Hz, 1H), 3.54-3.50 (m complex, 1H); 13C{1H} NMR (125 MHz, CDCI3) õ 161.2, 160.7, 150.6, 150.0, 144.4, 124.8, 119.2, 112.3, 110.7, 106.4, 100.9, 97.0, 78.8, 66.8, 56.4, 56.0, 55.6, 39.7; IR umax3392, 2919, 2850, 1620, 1495, 1464, 1418, 1385, 1267, 1192, 1145, 1132, 1082, 1026, 947, 846, 773 cm-1 ; HRMS (ESI, +ve) m / z [M+H]+ calculated for C18H18O5315.1227; Found 315.1227.

[0216] With the second synthesis developed here, pterocarpan compounds can be obtained enantioselectively for the (S,S) isomer with an overall yield of approximately 28%. Example 3 - Multigram synthesis according to Route 2 using Current Good Manufacturing Practices (c-GMP)

[0217] In the first step, a solution of the compounds 3,4-dimethoxyphenol (17.7 g, 115 mmol) and 2,4-dimethoxyphenylacetic acid (15.0 g, 76.6 mmol) in BF3*Et2O (56.8 ml, 460 mmol), magnetically stirred, contained in a 1000 ml round-bottom flask with an inlet septum equipped with a Teflon® stopcock, was washed with argon and stirred at 65 °C (oil bath) for 9 h. The resulting mixture, very dark and magnetically stirred, was cooled in an ice-water bath and then treated dropwise with DMF (118 ml), heated to 22 °C and, after 0.25 h at that temperature, heated to 50 °C (oil bath). Next, the magnetically stirred reaction mixture was treated with p-toluenesulfonyl chloride (43.7 g, 230 mmol), which was then added to the reaction mixture, which was then heated to 75 °C for 17.5 h. The cooled reaction mixture was diluted with ethyl acetate (800 mL) and treated with water (330 mL).The separated and heterogeneous lower phase was extracted with dichloromethane (3 x 200 mL). The combined organic phases were washed with water (2 x 330 mL) before being dried with sodium sulfate and then filtered through a cotton plug contained in a glass filter funnel. The filtrate was concentrated under reduced pressure (rotary evaporator) to give a nearly black oil. A solution of this oil in a minimum volume of ethyl acetate / petroleum ether / dichloromethane was added to the top of a chromatographic-grade silica gel pad contained in a flash chromatography column, and then the pad was eluted with dichloromethane. The concentration of the fractions... Relevant (Rf = 0.5 in 5:4 v / v petroleum ether / ethyl acetate) produced a brown oil that was dissolved in hot ethyl acetate and, after cooling the resulting solution, the compound 3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (14.1 g, 54%) was obtained as a light brown crystalline solid, mp = 218-219 °C. 1H NMR (500 MHz, CDCI3) 6.89 (s, 1H), 7.61 (s, 1H), 7.24 (d, J = 9.0 Hz, 1H), 6.87 (s, 1H), 6.56-6.54 (m-complex, 2H), 3.97(X) (s, 3H), 3.97(X) (s, 3H), 3.83 (s, 3H), 3.77 (s, 3H); 13C{1H} NMR (125 MHz, CDCl3) is 175.7, 161.2, 158.7, 154.3, 153.6, 152.5, 147.7, 132.4, 121.9, 118.1, 113.8, 105.2, 104.6, 99.7, 99.2, 56.5(3), 56.4(5), 55.9, 55.6.

[0218] In the second step, a magnetically stirred solution of 3-(2,4-dimethoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (13.0 g, 388 mmol) in acetonitrile (188 ml), contained in a 1000 ml round-bottom flask with an inlet septum fitted with a Teflon® stopcock, was washed with argon and cooled in an ice bath before being treated with AlCl3 (15.2 g, 114 mmol) and then with NaI (11.4 g, 76 mmol). The resulting mixture was heated to room temperature and, after 0.25 h, heated to 82 °C for 7.5 h. The cooled reaction mixture was placed in an ice bath and, while still stirring, slowly treated with ice and water (approximately 50 ml) and then diluted with water (200 ml) and then with sodium hydroxide (50 ml of a 2 M aqueous solution). The resulting mixture was extracted with ethyl acetate (1 x 600 mL) and the separated aqueous phase, containing a colloidal suspension, was filtered through a layer of diatomaceous earth contained in a sintered glass funnel.The filtrate was extracted with ethyl acetate (1 x 200 mL) and the combined organic phases were dried with sodium sulfate before filtration. The filtrate was then concentrated under reduced pressure in a rotary evaporator to provide a dark-colored solid. A solution of this solid in dichloromethane (100 mL) was filtered through a thin layer of chromatographic-grade silica gel contained in a flash chromatography column, and the filtrate was then concentrated under reduced pressure to provide a brown crystalline solid. This solid was dissolved in a minimum volume of hot ethyl acetate, and the resulting solution was cooled to room temperature. After 10 min, the solid thus formed was removed by filtration and then air-dried to provide the compound 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (7.03 g). 56%) as gold-colored crystalline masses, pf = 203-204 °C. 1 H RMN (500 MHz, CDCI3) õ 9.41 (s, 1 H), 8.06 (s, 1 H), 7.63 (s, 1 H), 7.07 (d, J = 8.5 Hz, 1 H), 6.94 (s, 1 H), 6.65 (d, J = 2.5 Hz, 2 H), 6.54 (dd, J = 8.5 e 2.5 Hz, 1 H), 4.01 (s, 3 H), 4.00 (s, 3 H), 3.82 (s, 3 H); RMN de 13C{1H} (125 MHz, CDCI3) õ 178.4, 162.1, 158.2, 155.5, 154.4, 148.5, 130.4, 124.6, 116.8, 113.2, 107.8, 104.6, 104.5, 99.3, 56.7, 56.6, 55.5.

[0219] In the third step, pure formic acid (9.3 ml, 247 mmol) was added dropwise to triethylamine (28.0 mL, 201 mmol), magnetically stirred, contained in a 500 ml round-bottom storage flask with a septum inlet fitted with a glass stopcock, which was washed with argon and cooled in an ice bath. The resulting mixture was allowed to warm to room temperature and, after 5 minutes, treated with a solution of RuCl(p-cymene)[(S,S)-Ts-DPEN] (1.085 g, 1.7 mmol, 0.08 molar equiv.) in DMSO (15 ml) and then with a suspension of the compound 3-(2-hydroxy-4-methoxyphenyl)-6,7-dimethoxy-4H-chromen-4-one (7.0 g, 21.3 mmol) in DMSO (35 ml). Next, the reaction mixture was heated to 45 °C for 24 h, then cooled to room temperature before being treated with hydrochloric acid (11 ml of a 37% aqueous solution).After a further 10 min, the reaction mixture was diluted with ammonium chloride (100 ml of a saturated aqueous solution), followed by water (100 ml), and the resulting mixture was then extracted with ethyl acetate (3 x 300 mL). The combined organic phases were washed with brine (1 x 50 ml), dried (Na2SO4), filtered and concentrated under reduced pressure (rotovap). The resulting dark brown oil was subjected to flash chromatography (silica, petroleum ether / ethyl acetate elution 3:1 v / v) to give, after concentration of the relevant fraction (Rf = 0.3), (+)-(6aS,11aS)-2,3,9-trimethoxy-6a,11a-dihydro-6H-benzofuro[3,2-c]chromene ((+)-PTC) (2.03 g, 31%) as a clear, light yellow oil, [a]D = +203° (c = 0.009, EtOH) {lit. [a]D = +203° (c = 1.3, EtOH)}. 1H NMR (500 MHz, CDCl3) õ 7.13 (d, J = 9.0 Hz, 1H), 6.98 (s, 1H), 6.48 (s, 1H), 6.46-6.44 (m complex, 2H), 5.49 (d, J = 7.0 Hz, 1H), 4.23 (dd, J = 10.5 and 4.5 Hz, 1H), 3.89 (s, 3H), 3.85 (s, 3H), 3.76 (s, 3H), 3.59 (t, J = 11.0 Hz, 1H), 3.55-3.51 (m complex, 1H); 13C{1H} NMR (125 MHz, CDCI3) õ 161.3, 160.7, 150.7, 150.1, 144.5, 124.9, 119.3, 112.4, 110.8, 106.5, 101.0, 97.0, 78.9, 66.8, 56.5, 56.0, 55.6, 39.8.

[0220] With the second synthesis developed here, pterocarpan compounds could be obtained enantioselectively for the (S,S) isomer with an overall yield of approximately 9%, on a multigram scale following Current Good Manufacturing Practices (c-GMP). Example 4 - Development of nanoformulations: liposome and immunoliposome Example 4.1 - Development of the liposome containing (+)-PTC

[0221] The liposome was developed with soy La-phosphatidylcholine (SPC), cholesterol (CHO), DSPE-PEG (2000) maleimide, and phosphatidylserine 16:0, with the molar proportions (%) defined experimentally and the amount of the (+)-PTC compound fixed at a total of 2 mg. The total amount of lipids, the ratio between SPC and CHO, and the addition of phosphatidylserine were evaluated to optimize the formulation to promote the highest encapsulation efficiency while maintaining acceptable size and polydispersity standards.

[0222] The lipid film hydration method described in “Diffusion of univalent ions across the lamellae of swollen phospholipids” by BANGHAM, Alec D.; STANDISH, Malcolm M.; WATKINS, Jeff C. (1965), incorporated here by reference, was used for liposome development. For the liposome loaded with (+)-PCT, the lipids and the compound were dissolved in 5 ml of chloroform. The chloroform was then removed, and a lipid film was formed using reduced pressure in a rotary evaporator for 20 minutes at 37 °C at 100 rpm. Immediately after, the lipid film was hydrated using a magnetic stirrer for 10 minutes at 100 rpm in an aqueous phase consisting of phosphate buffer (pH 7.4). Finally, the formulation was homogenized in a pointed sonicator (model 0125-OSonica) with a constant amplitude of 35% for 10 minutes on ice, to promote the formation of small unilamellar vesicles.

[0223] For the separation of the non-encapsulated compound, 0.45 µm filters were used, which retain lipophilic material, and the hydrophilic material is filtered out. Only the (+)-PTC compound, being extremely lipophilic, is retained by the filter. Example 4.2 - Development of the immunoliposome containing (+)-PTC

[0224] The immunoliposome was developed by functionalizing the (+)-PTC-encapsulated liposome produced in Example 4.1 using the anti-EGFR antibody (Cetuximab). This followed the method described in ELOY, Josimar O. et al. Anti-HER2 immunoliposomes for co-delivery of paclitaxel and rapamycin for breast cancer therapy. European journal of pharmaceuticals and biopharmaceutics, v. 115, p. 159-167, 2017, incorporated here by reference, with modifications in the proportions and quantities of material. First, for the functionalization of the liposome, the anti-EGFR antibody underwent a thiolation step; A solution containing 2 mg of antibody, 5 mM PBS / EDTA, pH 8.0, and 50 mM PBS / EDTA was prepared and adjusted with 1M NaOH to obtain pH 8. Then, Traut's reagent (2 mg / mL) was added in a 20:1 (w / w) ratio relative to the antibody. The solution was homogenized for 3 minutes and then incubated for 1 hour at 37 °C to promote the antibody thiolation process.

[0225] The thiolated antibody was separated from the free Traut reagent by gel filtration chromatography using a PD-10 column (Cytiva®) with PBS / EDTA5 mM eluent, pH 7. The fractions were collected and quantified by the BCA assay. After this step, the fractions were combined and added to the previously prepared liposome; the liposome-antibody solution was incubated overnight under constant agitation. The immunoliposome was then purified by gel filtration chromatography using a CL-4B column to prepare both the free antibody and the free (+)-PTC immunoliposomes, eluting with PBS buffer, pH 7.4. To define the fractions containing only the immunoliposome, encapsulated formulations with DiO₂, a fluorescent lipophilic marker, were prepared. During column separation, the fractions with the marker, which appear yellowish, can be visualized. Example 4.3 - Physicochemical characterization of liposomes and immunoliposomes

[0226] The particle size and polydispersity parameters of the formulations were analyzed using the Zetasizer Nano-NZ equipment (Malvern®), requiring the formulations to be diluted 10:1 (v / v) with ultrapure water; the surface charge was measured using the zeta potential using the same equipment.

[0227] To investigate the stability of the lyophilized formulations, liposome samples were added to 14 mg of sucrose, previously frozen at -80°C for 24 hours, and then lyophilized for another 24 hours. After the lyophilization process, the formulations were stored at 25°C and analyzed over a period of 15 days at 7-day intervals, measuring the parameters of size, zeta potential, and polydispersity. Example 4.4 - Evaluation of cellular uptake of the formulations

[0228] For evaluation of liposome uptake, total immunoliposome and purified immunoliposome were prepared without the (+)-PTC compound, but containing the lipid bilayer marker, DiO, encapsulated at a concentration of 0.1 mg / mL. The cells were plated at a concentration of 5x10 5 Cells were placed in 6-well plates and incubated for 24 hours at 37°C in an atmosphere containing 5% CO2. Then, the cells were washed with PBS, pH 7.4, and incubated with the diluted formulations for 15 minutes, 1 hour, and 3 hours. They were then washed again with PBS and trypsinized with 0.25% trypsin and inactivated with 1 ml of complete medium. After this step, the cells were centrifuged and resuspended in PBS. The samples were analyzed using the BD FACSVerse® cytometer. For all parameters evaluated, a total of 10,000 events were considered for each replicate of each sample. Example 4.5 - Evaluation of in vitro cytotoxicity of nanoformulations in the DU145 cell line.

[0229] The cytotoxic effect of treatment with free (+)-PTC, docetaxel, and the formulations on the inhibition of cell line growth was quantified using a colorimetric assay of a yellow tetrazolium salt (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide or MTT). The compounds were diluted in 100% DMSO to a stock concentration of 20 mM ((+)-PTC), 250 M (Docetaxel), and the formulations in PBS at pH 7.4 to a concentration of 800 pM, then stored at -80°C or 4°C (formulations). From the stock concentration, the compounds and formulations were diluted in DMEM or RPMI to develop the 14-point CRC. Cells were plated at specified concentrations (as per experimental condition), containing 50 pL of cells / well. Then, 24h after the Plating: 5 pL of positive control (DMSO 100%), negative control (DMSO 4%), (+)-PTC (80 pM-0.004 pM), formulations (80 pM - 0.004 pM), and docetaxel (1 pM - 0.00025 pM) to a final volume of 50 pL / well were added to each specific control and treatment group. After 72 hours, the MTT colorimetric assay was performed. For this, the plates were incubated with MTT for 4 h and the plates were read with the VictorNivo multimode plate reader (Perkin Elmer) using absorbance at 570 nanometers. The data were normalized relative to the negative control.

[0230] The absorbance vs. concentration graph was recorded, and its average inhibitory concentration capable of causing 50% of the maximum effect (IC50) and its respective 95% confidence intervals (95% CI) were determined using non-linear regression in GraphPad Prism Software version 6.0. Example 5.1 - Results Example 5.1.1 - Development of nanoformulations: liposome and immunoliposome

[0231] Tables 6 and 7 below summarize the formulations prepared for encapsulation of the (+)-PTC compound. In the developed formulations, 3 components were modified: the proportion of phospholipids (SPC), cholesterol (CHO), and total lipid. Both formulations 1 and 2 showed excellent size and polydispersity parameters (PDI), but with low encapsulation efficiency (EE%). Increasing the SPC concentration in formulation 2 provided a slight increase in EE%. Therefore, a new formulation (3) was developed using the proportions used in formulation 2 as a reference, but increasing the total lipid used. Formulation 3, containing a total of 80 mg of lipids, maintained excellent size parameters, PDI, zeta potential, as shown in Figures 10 and 11, and provided an EE% of 90%. Formulation 3 increased the EE%, but maintained a "drug loading" parameter similar to the other formulations.Thus, a new formulation (4) was developed following the same proportions as formulation 3, but with twice the amount of compound (4mg) and maintained the excellent parameters of size, PDI, zeta potential and EE% and with double the “drug loading” parameter going from a value of 2.25% to. 4.5% Formulation 3 was selected for further studies. Table 6 - Physicochemical characteristics of encapsulated formulations with (+)-PTC, part 1 Table 7 - Physicochemical characteristics of encapsulated formulations with (+)-PTC, part 2

[0232] The stability of formulation 3 was evaluated in its lyophilized form over a period of 15 days and compared with the non-lyophilized formulation, verifying the size, PDI, and zeta potential parameters. Figure 12 shows that the lyophilization process resulted in an increase in the size of the formulation, but without altering the other parameters. Regarding the lyophilized formulation, over the 15-day period there was minimal variation in the evaluated parameters, showing that the formulation in its lyophilized form was stable for a period of at least 15 days. Example 5.1.2 - Development of anti-EGFR immunoliposomes

[0233] For the development of the anti-EGFR immunoliposome loaded with (+)-PTC, formulation 3 was prepared and functionalized with the anti-EGFR antibody using the direct conjugation method, based on the thioether linkage between the thiol group of the antibody and the maleimide present on the liposome surface, which allows the formation of a stable and highly efficient bond, as illustrated in Figure 13. The chromatogram illustrated in Figure 14 corresponds to The anti-EGFR immunoliposome showed two distinct peaks: the first peak corresponds to the immunoliposome, where the compound with the highest molecular weight was eluted during gel filtration, and the second peak corresponds to the antibodies that were not conjugated. Thus, a conjugation efficiency of 47% was obtained using a total of 2 mg of antibodies in the formulation.

[0234] The anti-EGFR immunoliposome maintained similar physicochemical patterns to formulation 3, with a slight increase in the size of the formulation, which is explained by the presence of the antibody conjugated to the liposome (table 8). Table 8 - Characterization of anti-EGFR liposome and immunoliposome formulations

[0235] The structure and integrity of the monoclonal antibody conjugated to the liposome was verified using the Western blot technique. The antibody present in the immunoliposome showed the same structural profile. Example 5.1.3 - Evaluation of cellular uptake of DU145 cell line formulations

[0236] Flow cytometry studies were performed on the DU145 cell line, with the two groups of formulations tested (liposome and purified immunoliposome). Treatment times of 15 minutes, 1 hour, and 3 hours were chosen for analysis. The fluorophore propidium iodide, a nucleic acid marker, was used to detect cell death. The fluorophore Dio, a marker of lipid bilayers encapsulated in liposomes and immunoliposomes, was used to visualize cell uptake. As shown in Figure 15, after 15 minutes of treatment, only the group treated with purified immunoliposome showed partial internalization of the formulation, and after 3 hours, more than 90% of the formulation was internalized. The liposome showed greater internalization after 3 hours, with a total of 75% cell internalization. This result demonstrates that functionalization with the anti-EGFR antibody promoted greater internalization. Rapid formulation. However, after 3 hours of incubation, both formulations showed a high rate of cellular internalization. There is a statistically significant difference (p<0.05) between the liposome and immunoliposome formulations at the evaluated times defined by the one-way AN OVA test. This fact is indicated by the use of asterisks in the bar graph of Figure 15, where ** p < 0.01 and **** p < 0.0001.

[0237] An indirect ELISA experiment was performed to evaluate the anti-EGFR binding capacity of the immunoliposome. As illustrated in Figure 16, it was observed that the liposome does not exhibit any binding to EGFR; however, both immunoliposome formulations show EGFR binding, characterized by increased absorbance when compared to the negative control and the liposome-treated group. Furthermore, the immunoliposomes exhibit the same binding capacity when compared to the reference antibody (Cetuximab). Example 5.1.4 - Evaluation of the cytotoxicity of the formulations in the DU145 cell line

[0238] The formulations were evaluated to determine their cytotoxic potential against the DU145 cell line after 72 hours of treatment, as shown in Figure 17. The formulations presented a similar concentration-response curve to the (+)-PTC compound.

[0239] Table 9 presents the IC50 values ​​obtained for (+)-PTC, immunoliposome, liposome, blank liposome, cetuximab, and docetaxel, being, respectively, 1.34 pM, 2.10 pM, >80 pM, 1.6 pM, >500 pg / mL, and 0.003 pM. Table 9 - IC50 of the formulations and the compounds (+)-PTC, Docetaxel, and Cetuximab evaluated in the DU145 cell line, with their respective confidence intervals. Example 5.2 - Discussions and Conclusions Example 5.2.1 - Development of nanoformulations

[0240] Cytotoxicity analysis of the developed nanoformulations demonstrated that they do not alter the cytotoxic effect of the (+)-PTC compound against the DU145 cell line, exhibiting similar IC50 values. Furthermore, in this model, free drugs can be transported directly into cells by passive diffusion, without a drug release process. Therefore, advantages of using the nanoformulations of the present invention can be observed, such as accumulation in the tumor mass, longer drug half-life, and greater specificity for tumor cells.

[0241] In vitro antitumor evaluations showed quite similar results, with no improvement in cytotoxic effect. On the other hand, in vivo antitumor studies demonstrated that the nanoformulations exhibit a significantly better antitumor effect than the free drug.

[0242] Among the formulations tested, those that presented the best physicochemical parameters and encapsulation efficiency were formulations 3 and 4, formed by SPC:CHO:DSPE-PEG-MAL:phosphatidylserine; 65:30:3:2, respectively. The proportion and constituents of formulation 3 are similar to the others tested, however, for this formulation a total of 80mg of lipids was used, while in the others 50mg was used. This increase promoted an improvement in encapsulation efficiency from 70% to 90%. For formulation 4, the amount of active compound was doubled and the advantageous physicochemical parameters of formulation 3 were preserved.

[0243] Physicochemical analyses of liposomes and immunoliposomes showed sizes of 80 nm and 90 nm, respectively, followed by a PDI of 0.2 and a zeta potential of -30 mV. These values ​​position the formulations as candidates for use in cancer treatment. Studies demonstrate that particle size significantly affects several pharmacokinetic parameters, including tissue extravasation, diffusion, hepatic uptake, renal excretion, and clearance rate after injection. Consequently, the ideal size range is between 80 and 150 nm and the zeta potential range is between -10 mV and -30 mV.

[0244] Analyzing the stability of nanoformulations is an important factor in nanoformulation, aiming to ensure the durability of nanoparticles over a prolonged period—a fundamental aspect for their therapeutic viability. Consequently, there is a pressing need for a strategy to maintain the efficacy of nanoparticles over time.

[0245] Lyophilization, also known as freeze-drying, has emerged as an important solution to meet this challenge.

[0246] The freeze-drying process maintained the physicochemical parameters of the nanoformulation, with a slight increase in size, but keeping it within the permitted range of values.

[0247] Regarding the development of the immunoliposome, the formulation that presented the best physicochemical parameters and encapsulation efficiency was chosen for the functionalization step with the anti-EGFR antibody. The strategy used is based on the interaction between the maleimide group present in the liposomes and the thiolated antibody, promoting a covalent bond between the liposome and the antibody.

[0248] For the immunoliposome of the present invention, the functionalization efficiency was 49%. The functionalization step did not alter the primary structure or the binding capacity of the antibody. The indirect ELISA assay indicates that conjugation of cetuximab to the liposome was satisfactory, maintaining its conformation suitable for effective binding to EGFR. This result reinforces the findings of cell uptake, demonstrating that post-conjugation of the antibody to the liposome preserves its functionality, leading to greater cell uptake compared to the non-functionalized liposome, at all times evaluated in the DU145 cell line.

[0249] Analysis of immunoliposome binding using ELISA demonstrated that the immunoliposome maintained a similar binding capacity to the free antibody. This result was supported by cellular uptake data, in which the immunoliposome promoted a significant increase in cellular uptake when compared to the liposome.

[0250] Analysis of the cytotoxicity of the developed nanoformulations demonstrated that the nanoformulations do not alter the cytotoxic effect of the lead (+)-PTC compound against the DU145 cell line, showing similar IC50 values.

[0251] Thus, the development of the nanoformulations was satisfactory, as it generated functional liposomes and immunoliposomes within acceptable parameters for their use in cancer treatment. Furthermore, the expression system of the biosimilar prototype was adequate, since the antibody presented structural and functional patterns similar to the reference antibody.

Claims

CLAIMS 1. Production process of reduced hydroxy-substituted isoflavone of formula (II): characterized by comprising the asymmetric hydrogen transfer step by contact of hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya ruthenium (S,S) complex in the presence of a hydrogen donor, wherein the process is enantioselective for the reduced hydroxy-substituted isoflavone (S,S) of formula (I la): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

2. Production process of reduced hydroxy-substituted isoflavone with formula (Ha): characterized by comprising the asymmetric hydrogen transfer step by contact of hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya ruthenium (S,S) complex in the presence of a hydrogen donor, wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy.

3. Process for producing reduced hydroxy-substituted isoflavones according to claim 1 or 2, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen, hydroxyl, Ci-Ce alkoxy or Ci-Ce haloalkoxy; R5, R6, R7, and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

4. Process for producing reduced hydroxy-substituted isoflavones according to any one of claims 1 to 3, characterized in that the hydrogen donor is selected from salts of carboxylic acids, carboxylic acids, amines, or a combination thereof.

5. A process for producing reduced hydroxy-substituted isoflavones according to claim 4, characterized in that the hydrogen source is a carboxylic acid salt selected from an alkali metal C1-C1 carboxylic acid salt, preferably lithium, sodium or potassium.

6. Process for producing reduced hydroxy-substituted isoflavone according to any one of claims 1 to 5, characterized in that the Noyori-lkariya Ruthenium (S,S) complex is a Noyori-lkariya Ruthenium (S,S) complex of formula (VII): in what M is ruthenium; r|6-Ar is an M-linked hapticity arene ligand selected from optionally substituted arene with Ci-Ce alkyl or Ci-Ce alkoxy; Ph stands for phenyl; X is a halogen; and R13 is arylsulfonyl.

7. Process for producing reduced hydroxy-substituted isoflavones according to any one of claims 1 to 6, characterized in that the asymmetric hydrogen transfer step occurs over a period of 2 hours to 24 hours.

8. Process for producing reduced hydroxy-substituted isoflavones according to any one of claims 1 to 7, characterized in that the asymmetric hydrogen transfer step occurs in a temperature range of 25 °C to 45 °C.

9. Process for producing reduced hydroxy-substituted isoflavone according to any one of claims 1 to 8, characterized in that the hydroxy-substituted isoflavone of formula (III) is produced from the selective dealkylation of the alkyloxy-substituted isoflavone of formula (IV): in which: Each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C6 alkoxy, or C1-C1 haloalkoxy; and R9 is selected from C1-C1 alkyl.

10. Process for producing reduced hydroxy-substituted isoflavones according to claim 9, characterized in that selective dealkylation occurs in the presence of a Lewis acid, preferably boron trichloride, and a polar aprotic solvent, preferably dichloromethane.

11. Process for producing reduced hydroxy-substituted isoflavones according to claim 9 or 10, characterized in that selective dealkylation occurs over a period of 0.5 to 24 hours.

12. Process for producing reduced hydroxy-substituted isoflavone according to any one of claims 9 to 11, characterized in that the 2'-alkyloxy-substituted isoflavone of formula (IV) is produced from the coupling of the 4H-chromen-4-one substituted of formula (V): in which: Each R1, R2, R3, and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy; and X is a halogen; with the substituted phenyl of formula (VI): in which: Each R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy. R9 is selected from Ci-Ce alkyl, and R10 is a leaving group B(OH)2.

13. Process for producing reduced hydroxy-substituted isoflavones according to claim 12, characterized in that the coupling is carried out in the presence of a palladium catalyst.

14. Process for producing reduced hydroxy-substituted isoflavones according to claim 13, characterized in that the coupling is carried out in the presence of an inorganic Lewis base, preferably carbonate, phosphate or hydroxide, and polyethylene glycol solvent.

15. Process for producing reduced hydroxy-substituted isoflavones according to any one of claims 12 to 14, characterized in that the coupling occurs in a temperature range of 25 to 50°C.

16. Process for producing reduced hydroxy-substituted isoflavones according to any one of claims 12 to 15, characterized in that coupling occurs between 1 hour and 24 hours.

17. Process for producing reduced hydroxy-substituted isoflavone according to any one of claims 12 to 16, characterized in that the substituted 4H-chromen-4-one of formula (V) is produced from the one-pot two-step reaction of acetophenone of formula (VIII): in which: Each R1, R2, R3, and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy; with DMF-DMA to form an enaminone intermediate, and addition of molecular halogen and pyridine to form the substituted 4H-chromen-4-one of formula (V).

18. Process for producing reduced hydroxy-substituted isoflavone according to claim 17, characterized in that the acetophenone of formula (VIII) is produced from the acylation of the phenol of formula (IX): wherein each R1, R2, R3 and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy; with acetic anhydride in the presence of a Lewis acid.

19. Production process of pterocarpan of formula (I): characterized by comprising the acid cyclization step of the reduced hydroxy substituted isoflavone of formula (II): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

20. Process for producing pterocarpan according to claim 19, characterized in that the hydroxy-substituted reduced isoflavone of formula (II) is obtained by the process as defined in any one of claims 1 to 18.

21. Production process of pterocarpane according to claim 19, characterized in that the reduced hydroxy-substituted isoflavone is composed mainly of the (S,S) isomer with formula (Ha): and pterocarpane is composed mainly of the (S,S) isomer with formula (1a) wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

22. Production process of pterocarpan of formula (la): characterized by comprising the acid cyclization step of the reduced hydroxy-substituted isoflavone with the formula (Ha): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, or C1-C6 haloalkoxy.

23. A process for producing pterocarpane according to any one of claims 19 to 22, characterized in that the cyclization in an acidic medium is carried out in the presence of a strong practical acid, preferably hydrochloric or sulfuric acid, and an aliphatic alcohol and ester solvent system, preferably ethanol with ethyl acetate.

24. Process for producing pterocarpan according to any one of claims 19 to 23, characterized in that cyclization occurs at room temperature.

25. A process for producing pterocarpane according to any one of claims 19 to 24, characterized in that cycling occurs over a period of 5 minutes to 1 hour.

26. Production process of pterocarpan of formula (I): characterized by comprising the reductive cyclization step of the hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya ruthenium (S,S) complex in the presence of a hydrogen donor, wherein the process is enantioselective for the pterocarpane (S,S) of formula (la): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

27. Production process of pterocarpan of formula (la): characterized by comprising the enantioselective reductive cyclization step of the hydroxy substituted isoflavone of formula (III): with a Noyori-lkariya ruthenium (S,S) complex in the presence of a hydrogen donor, wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

28. Process for producing pterocarpan according to claim 26 or 27, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen, hydroxyl, Ci-Ce alkoxy or Ci-Ce haloalkoxy; R5, R6, R7, and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

29. A process for producing pterocarpane according to any one of claims 26 to 28, characterized in that the hydrogen donor is selected from salts of carboxylic acids, carboxylic acids, amines, or a combination thereof.

30. A process for producing pterocarpane according to claim 29, characterized in that the hydrogen donor is a mixture of carboxylic acid and amine, preferably a mixture of formic acid and triethylamine.

31. Process for producing pterocarpane according to any one of claims 26 to 30, characterized in that the complex Noyori-lkariya of Ruthenium (S,S) is a Noyori-lkariya of Ruthenium (S,S) complex of formula (VII): in what M is ruthenium; r|6-Ar is a 6-hapticity arene ligand linked to M selected from arene optionally substituted with Ci-Ce alkyl or Ci-Ce alkoxy; Ph stands for phenyl; X is a halogen; and R13 is arylsulfonyl.

32. A process for producing pterocarpane according to any one of claims 26 to 31, characterized in that the reductive cycling step takes place over a period of 1 hour to 48 hours.

33. A process for producing pterocarpane according to any one of claims 26 to 32, characterized in that the reductive cyclization step occurs in a temperature range of 0 °C to 60 °C.

34. Process for producing pterocarpane according to any one of claims 26 to 33, characterized in that the hydroxy-substituted isoflavone of formula (III) is produced from the selective dealkylation of the alkyloxy-substituted isoflavone of formula (IV): in which: Each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 haloalkoxy; and R9 is selected from C1-C1 alkyl.

35. A process for producing pterocarpane according to claim 34, characterized in that selective dealkylation occurs in the presence of a Lewis acid, preferably an aluminum halide, a nucleophile source, preferably an alkali metal halide, and a polar aprotic solvent, preferably acetonitrile.

36. Process for producing pterocarpan according to claim 34 or 35, characterized in that selective dealkylation occurs over a period of 0.5 to 24 hours.

37. Process for producing pterocarpane according to any one of claims 34 to 36, characterized in that the alkyloxy substituted isoflavone of formula (IV) is produced from the one-pot two-step reaction of the phenol of formula (IX): wherein each R1, R2, R3 and R4 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy; with aryl acetic acid of formula (X): wherein each R5, R6, R7 and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy or C1-C1 haloalkoxy, and R9 is selected from C1-C1 alkyl, using a Lewis acid for acylation with the formation of a ketone intermediate in a first step and subsequent formylation / cyclization / dehydration with a polar aprotic solvent and dehydrating agent in a second step.

38. A process for producing pterocarpane according to claim 37, characterized in that the Lewis acid is selected from metal chloride or diethyl borotrifluoride, preferably diethyl borotrifluoride.

39. A process for producing pterocarpane according to claim 37 or 38, characterized in that the polar aprotic solvent is selected from dichloromethane, acetonitrile, dimethylformamide, dimethyl sulfoxide, and tetrahydrofuran, preferably dimethylformamide.

40. Process for producing pterocarpane according to any one of claims 37 to 39, characterized in that the dehydrating agent is selected from p-toluenesulfonyl chloride or phosphorus oxychloride.

41. A process for producing pterocarpane according to any one of claims 37 to 40, characterized in that acylation occurs in a temperature range of 0 °C to 80 °C, for a period of 1 to 24 hours.

42. A process for producing pterocarpane according to any one of claims 37 to 41, characterized in that formylation / cyclization / dehydration occurs in a temperature range of 20 °C to 100 °C, for a period of 1 to 48 hours.

43. Pterocarpan compound, characterized by having formula (I) wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

44. Pterocarpane compound, according to claim 43, characterized by having formula (1a) wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C1 alkyl, C1-C1 haloalkyl, C1-C1 alkoxy, or C1-C1 haloalkoxy.

45. Pterocarpane compound, according to claim 43 or 44, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen, hydroxyl, C1-C1 alkoxy or C1-C1 haloalkoxy; and R5, R6, R7 and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

46. ​​Pterocarpane compound, according to claim 43 or 44, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen or Ci-Ce alkoxy or Ci-C6 haloalkoxy; R5, R6, and R8 are hydrogen; and R7 is Ci-Ce alkoxy 47. Pterocarpane compound, according to claim 43 or 44, characterized in that: R1 and R4 are hydrogen. R2 and R3 are Ci-Ce alkoxy compounds; and R5, R6, R7, and R8 are selected independently of hydrogen or Ci-Ce alkoxy.

48. Hydroxy-substituted reduced isoflavone, characterized by having formula (II): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy or C1-C6 haloalkoxy.

49. Hydroxy-substituted reduced isoflavone, according to claim 48, characterized by having the formula (Ha): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy or C1-C6 haloalkoxy.

50. Hydroxy-substituted reduced isoflavone, according to claim 48 or 49, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen, hydroxyl, C1-C1 alkoxy or C1-C1 haloalkoxy; and R5, R6, R7 and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

51. Hydroxy-substituted reduced isoflavone, according to claim 48 or 49, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen or Ci-Ce alkoxy or Ci-C6 haloalkoxy; R5, R6, and R8 are hydrogen; and R7 is Ci-Ce alkoxy 52. Hydroxy-substituted reduced isoflavone, according to claim 48 or 49, characterized in that: R1 and R4 are hydrogen. R2 and R3 are Ci-Ce alkoxy compounds; and R5, R6, R7, and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

53. Hydroxy-substituted isoflavone, characterized by having formula (III): wherein each R1, R2, R3, R4, R5, R6, R7, and R8 is independently selected from hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy or C1-C6 haloalkoxy.

54. Hydroxy-substituted isoflavone, according to claim 53, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen, hydroxyl, C1-C1 alkoxy or C1-C1 haloalkoxy; and R5, R6, R7 and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

55. Hydroxy-substituted isoflavone, according to claim 53, characterized in that: R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen or Ci-Ce alkoxy or Ci-C6 haloalkoxy; R5, R6, and R8 are hydrogen; and R7 is Ci-Ce alkoxy 56. Hydroxy-substituted isoflavone, according to claim 53, characterized in that: R1 and R4 are hydrogen. R2 and R3 are Ci-Ce alkoxy compounds; and R5, R6, R7, and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

57. Liposome, characterized by the fact that it comprises lipids and a pterocarpan compound.

58. Liposome, according to claim 57, characterized in that the pterocarpan compound is the pterocarpan compound of formula (I) as defined in any one of claims 43 to 47.

59. Liposome, according to claim 57 or 58, characterized in that the pterocarpan compound of formula (I) is in its dextrorotatory, levorotatory form or in a stereoisomeric or racemic mixture thereof.

60. Liposome according to claim 58, characterized in that, in the pterocarpan compound of formula (I): R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen, hydroxyl, C1-C1 alkoxy or C1-C1 haloalkoxy; and R5, R6, R7 and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

61. Liposome according to claim 57, characterized in that, in the pterocarpan compound of formula (I): R1, R2 and R4 are independently selected from hydrogen or Ci-Ce alkoxy; R3 is selected from hydrogen or Ci-Ce alkoxy or Ci-C6 haloalkoxy; R5, R6, and R8 are hydrogen; and R7 is Ci-Ce alkoxy 62. Liposome according to claim 57, characterized in that, in the pterocarpan compound of formula (I): R1 and R4 are hydrogen. R2 and R3 are Ci-Ce alkoxy compounds; and R5, R6, R7, and R8 are independently selected from hydrogen or Ci-Ce alkoxy.

63. Liposome, according to any one of claims 57 to 62, characterized in that the lipids are selected from phospholipids, cholesterol or a combination thereof.

64. Liposome, according to claim 63, characterized in that the phospholipid is selected from phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidylinositol, distearoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, dileoylphosphatidylethanolamine, or a combination thereof.

65. Liposome, according to any one of claims 57 to 64, characterized in that it further comprises a stealth agent, preferably selected from polyethylene glycol, polysarcin, polycarboxybetaine or a combination thereof.

66. Liposome, according to any one of claims 57 to 64, characterized in that it further comprises a conjugating agent, preferably selected from maleimide, iodoacetamide, N-hydroxysuccinimide, carbodiimide, or a combination thereof.

67. Liposome, according to any one of claims 65 to 66, characterized in that the liposome comprises a stealth agent and a conjugating agent present in a heterobifunctional conjugating agent comprising an anchoring agent, the stealth agent and the conjugating agent, preferably DSPE-PEG-MAL.

68. Liposome, according to any one of claims 57 to 67, characterized in that it comprises from 2% to 10% by weight of pterocarpan compound, based on the total weight of the liposome.

69. Liposome, according to any one of claims 57 to 68, characterized in that it comprises from 45 mol% to 80 mol% by weight of phospholipids, based on total moles of lipids.

70. Liposome, according to any one of claims 62 to 69, characterized in that it comprises from 10 mol% to 45 mol% of cholesterol, based on total moles of lipids.

71. Liposome, according to any one of claims 57 to 70, characterized in that it has a drug-to-lipid ratio of 1 to 10%.

72. Liposome, according to any one of claims 57 to 71, characterized in that it is an immunoliposome functionalized with a monoclonal antibody.

73. Liposome production process, as defined in any one of claims 57 to 72, characterized by comprising the steps of: a) dilution of the supporting phase material in a volatile organic solvent b) formation of a lipid film c) evaporation of the volatile organic solvent d) solubilization of the lipid film in the aqueous phase e) fragmentation to obtain single-layer nanoparticles.

74. Liposome production process according to claim 73, characterized by further comprising step f) of solution purification.

75. Liposome production process according to claim 73 or 74, characterized by further comprising step g) of conjugating the monoclonal antibody to the liposome.

76. Pharmaceutical composition, characterized by comprising the pterocarpan compound as defined in any one of claims 43 to 47, or the liposome as defined in any one of claims 57 to 72.

77. Use of the pterocarpan compound, as defined in any one of claims 43 to 47, of the liposome as defined in any one of claims 57 to 72, or of the pharmaceutical composition as defined in claim 76, characterized in that it is in the manufacture of a medicament for treating cancer.

78. Use according to claim 77 characterized by the cancer being selected from the group consisting of breast, prostate, leukemia, myelodysplastic syndrome, glioma, colon, lung and ovarian cancer.

79. Use, according to claim 78, characterized by the cancer being prostate cancer.

80. Use, according to any of claims 77 to 79, characterized in that the medication is administered intravenously or intraperitoneally.

81. Treatment method, characterized in that it comprises administering the pterocarpan compound, as defined in any one of claims 43 to 47, the liposome as defined in any one of claims 57 to 72, or the pharmaceutical composition as defined in claim 76 to an individual with cancer.

82. Treatment method according to claim 81, characterized in that the cancer is selected from the group consisting of breast, prostate, leukemia, myelodysplastic syndrome, glioma, colon, lung and ovarian cancer.

83. Treatment method according to claim 82, characterized in that the cancer is selected from the group consisting of prostate cancer.

84. Treatment method, according to any one of claims 81 to 83, characterized by administration being intravenous or intraperitoneal.

Images