MXPA00008737A

MXPA00008737A - Steroid derived antibiotics

Info

Publication number: MXPA00008737A
Application number: MXPA/A/2000/008737A
Authority: MX
Inventors: Paul B Savage; Chunhong Li
Original assignee: Brigham Young University
Filing date: 2000-09-06
Publication date: 2002-07-25

Abstract

A series of novel steroid derivatives are described. The steroid derivatives are antibacterial agents. The steroid derivatives also act to sensitize bacteria to other antibiotics including erythromicin and novobiocin.

Description

ANTI-BIÓTICOS DERIVADOS DE ESTEROI DES BACKGROUND OF THE INVENTION The invention relates to novel steroid derivatives and salts thereof and the processes and intermediates for their preparation. Some compounds that strongly associate with the outer membrane of Gram-negative bacteria are known to break down the outer membrane and increase permeability. The increased permeability may lead directly to cell death or may increase the susceptibility of Gram-negative bacteria to other antibiotics. The most studied of this type of compound are polymyxin antibiotics. For an example of a study involving the binding of polymyxin B to the primary constituent of the outer membrane of Gram-negative bacteria (lipid A) see: D.C. Morrison and D. M. Jacobs, Binding of Polymixin B to the Lipid to Portion of Bacterial Lipopolysaccharides (Binding of Polymembrane B to the Lipid, a Portion of Bacterial Lipopolysaccharides), Immunochemistry 1 976, vol. 1 3, 81 3-81 9. For an example of a study involving the binding of a polymyxin derivative to Gram-negative bacteria see: M. Vaara and P. Viljanen, Binding of Polymyxin B Nonapeptide to Gram-negative Bacteria, (Nonapeptide binding of polymyxin B to Gram-negative bacteria), Antimicrobial Agents and Chemoterapy, 1 985, vol. 27, 548-554. Membranes of Gram-negative bacteria are semipermeable molecular "sieves", which restrict the access of antibiotics and host defense molecules to their targets within the bacterial cell. In this way, the cations and polycations that access the system of self-promoted uptake are, by virtue of their ability to interact with and break the outer membrane permeability barrier, capable of increasing the susceptibility of pathogenic Gram-negative bacteria to antibiotics. and host defense molecules. Hancock and Wong demonstrated that a wide range of peptides could overcome the permeability barrier and coined the name "permeabilizing" to describe them (Hancock and Wong, Antimicrob Agents Chemother., 26:48, 1884).

BRIEF DESCRIPTION OF THE INVENTION The present invention features compounds of the formula I wherein: the fused rings A, B, C and D are independently saturated or fully or partially unsaturated; and Ri to R4, R6, R, R11, R? 2, R15, R1 and R17 is each independently selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl, (C1-C10) alkyloxy- (C1-) C10) alkyl, a substituted or unsubstituted (C1-C10) aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo, a linker group attached to a second steroid, a (C1-C10) substituted or unsubstituted aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O, H2N -HC (Q5) -C (O) -N (H) -, (C1-C10), azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O-, ( C1-C10) guanidinoalkyl oxy, and (C1-C10) guanidinoalkyl carboxy, where Q5 is a side chain of any amino acid, PG is an amino protecting group, and R5, R8, R9, Rio, R13 and R4 are each independently: deleted when one of the fused rings A, B, C or D is unsaturated in order to complete the valency of the atom carbon at that site, or selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl , a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo, a linker group attached to a second steroid, a (C1- C10) substituted or unsubstituted aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O-, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10) azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG- HN-C (Q5) -C (O) -O, (C1-C10) guanidinoalkyloxy and (C1-C10) guanidinoalkylcarboxy, where Q5 is a side chain of any amino acid, PG is an amino protecting group, and provided that at least two of R-α to R1 are independently selected from the group consisting of a (C 1 -C 1 0) substituted or unsubstituted aminoalkyloxy, a (C 1 -C 1 0) substituted or unsubstituted aminoalkylcarboxy, a substituted or unsubstituted (C 1 -C 1 0) aminoalkylaminocarbonyl, H 2 N -CH (Q 5) -C (O) -O-, H 2 N-HC (Q 5) -C (O) -N ( H) -, (C 1 -C 10) azidoalkyloxy, (C 1 -C 10) cyanoalkyloxy, PG-HN-C (Q 5) -C (O) -O-, (C 1 -C 10) guanidinoalkyloxy and ( C 1 -C 1 0) guanidinoalkylcarboxy; or a pharmaceutically acceptable salt thereof. The term "fused ring" used herein may be heterocyclic or carbocyclic, preferably. The term "saturated" as used herein, refers to the fused ring of formula I having each atom in the ring fused, either hydrogenated or substituted so that the valence of each atom is full. The term "unsaturated" as used herein, refers to the fused ring of formula I, wherein the valence of each atom of the fused ring may not be filled with hydrogen or another substituent. For example, the adjacent carbon atoms in the fused ring may be double-bonded to one another. The unsaturation may also include suppressing at least one of the following pairs and completing the valency of the ring carbon atoms in these suppressed positions with a double bond; such as, R5 and R9; R8 and R1 0; and R13 and R1.

The term "unsubstituted" used herein, refers to a portion having each hydrogenated atom, so that the valence of each atom is full. The term "halo" used herein, refers to a halogen atom, such as, fluorine, chlorine, bromine or iodine. Examples of amino acid side chains include, but are not limited to H (glycine), methyl (alanine), - (CH2- (C = O) -NH2 (asparagine), -CH2-SH (cysteine), and -CH ( OH) CH3 (threonine) An alkyl group is a linear or branched hydrocarbon which can be substituted or unsubstituted Examples of branched alkyl groups include isopropyl, sec-butyl, isobutyl, tert-butyl, sec-pentyl, isopentyl, ter -pentyl, isohexyl Substituted alkyl groups may have one, two, three or more substituents, which may be the same or different, each replacing a hydrogen atom Substituents are halogen (e.g., F, Cl, Br e I), hydroxyl, protected hydroxyl, amino, protected amino, carboxy, protected carboxy, cyano, methylsulfonylamino, alkoxy, acyloxy, nitro and lower haloalkyl The term "substituted" used herein refers to portions having one, two, three or more substituents, which may be the same or different, each one placing a hydrogen atom. Examples of substituents include, but are not limited to halogen (e.g., F, Cl, Br and I), hydroxyl, protected hydroxyl, amino, protected amino, carboxy, protected carboxy, cyano, methylsulfonylamino, alkoxy, alkyl, acryl, acyloxy, nitro and lower haloalkyl.

An aryl group is an aromatic ring of C6.20, wherein the ring is made of carbon atoms (for example, aryl groups of C6, C6, 10). Examples of haloalkyl include fluoromethyl, dichloromethyl, trifluoromethyl, 1,1-difluoroethyl and 2,2-dibromoethyl. A linker group is a divalent moiety used to link a compound of formula to another steroid, for example, a second compound of formula I. An example of a linker group is (C 1 -C 1 0) alkyloxy- (C 1 -C 0) alkyl. Numerous amino-protecting groups are well known in the art. In general, the protective group species is not critical, as long as it is stable for the conditions of any reaction or subsequent reactions in other positions of the compound, and can be removed at the appropriate point without adversely affecting the rest of the molecule. In addition, one protective group can be replaced by another after the substantive synthetic transformations are complete. Clearly, where a compound differs from a compound described herein only in that one or more protecting groups of the described compound have been substituted with a different protecting group, that compound is within the invention. Examples and additional conditions are found in T.W. Greene, Protective Groups in Organic Chemistry, (1st ed., 1 981, 2nd ed., 1991). The present invention also includes methods for synthesizing compounds of formula I, wherein at least two of R- and R14 are independently selected from the group consisting of a substituted or unsubstituted (C1-C10) aminoalkyloxy. The method includes the step of contacting a compound of formula IV, where at least two of R, through R1 are hydroxyl, and the remaining portions in the fused rings A, B, C and D are defined by formula I, with an electrophile to produce an alkyl ether compound of formula IV , wherein at least two of Ri to R1 are (C 1 -C 1 0) alkyloxy. The alkyl ether compounds are converted to an amino precursor compound, wherein at least two of R ^ to R14 are independently selected from the group consisting of (C 1 -C 1 0) azidoalkyloxy and (C 1 -C 1 0) cyanoalkyloxy and the amino precursor compound is reduced to form a compound of formula I. Electrophiles used in the method include but are not limited to 2- (2-bromoethyl) -1,3-dioxolane, 2-iodoacetamide, 2-chloroacetamide,? / - (2-bromoethyl) phthalimide,? / - (3-bromopropyl) phthalimide and allyl bromide. The preferred electrophile is allyl bromide. The invention also includes a method for producing a compound of formula I, wherein at least two of R ^ to R14 are (C1-C10) guanidoalkyloxy. The method includes contacting a compound of formula IV, wherein at least two of R, and Ri are hydroxyl, with an electrophile to produce an alkyl ether compound of formula IV, wherein at least two of R to R? 4 are ( C1-C10) alkyloxy. The allyl ether compound is converted to an amino precursor compound, wherein at least two of R to R 14 are (C 1 -C 1 0) aminoalkyloxy. The aminoalkyl ether compound is contacted with a guanidino producing electrophile to form a compound of formula I. The term "guanidino producing electrophile" used herein, refers to an electrophile used to produce a guanidino compound of formula I. An example of a guanidino-producing electrophile is HSO3-C / NH) -NH2. The invention also includes a method for producing a compound of formula I, wherein at least two of R ^ to R1 are H2N-HC (Q5) -C (O) -O- and Q5 is the side chain of any amino acid. The method includes the step of contacting a compound of formula IV, wherein at least two of R, to R14 are hydroxyl, with a protected amino acid to produce a protected amino acid compound of formula IV, wherein at least two of at least of R ^ a R are PG-HN-HC (Q5) -C (O) -Oy Q5 is the side chain of any amino acid and PG It is an amino protecting group. The protecting group of the protected amino acid compound is removed to form a compound of formula I. The present invention also includes pharmaceutical compositions of matter that are useful as antibacterial agents, sensitizers of bacteria to other antibiotics and bacterial membrane breakers.

The pharmaceutical compositions can be used to treat humans and animals that have a bacterial infection. The pharmaceutical compositions may include an effective amount of the steroid derivative alone or in combination with other antibacterial agents. Without wishing to join a particular theory, the steroid derivatives act as bacteriostatic and bactericidal agents by binding the outer cell membrane of the bacterium. The interaction between the steroid derivatives and the bacterial membrane breaks the integrity of the cell membrane and results in the death of the bacterial cell. In addition, the compounds of the present invention also act to sensitize the bacteria to other antibiotics. At concentrations of the steroid derivatives below the corresponding bacteriostatic concentration, the derivatives cause the bacteria to become more susceptible to other antibiotics by increasing the permeability of the outer membrane of the bacteria. Measurements used to quantify the effects of steroid derivatives on bacteria include: measurement of minimum inhibitory concentrations (MICs), measurement of minimal bactericidal concentrations (MBCs), and the ability of steroid derivatives to lower the M ICs of other antibiotics, for example, erythromycin and novobiocin. A person of experience will recognize that the compounds described herein retain certain stereochemical and electronic characteristics found on steroids. The term "same configuration", as used herein, refers to substituents on the fused steroid having the same stereochemical orientation. For example, the substituents R3, R7 and R? 2 are all β-substituted or a-substituted. The configuration of the substituted portions R3, R and R? 2 in C3, C7 and C12 may be important for interaction with the cell membrane. In another aspect, the invention features various methods for using the compounds described above. For example, an effective amount of an anti-microbial composition comprising such a compound is administered to a host (including a human host) to treat a microbial infection. The compound itself can provide the anti-microbial effect, in which case the amount of the compound administered is sufficient to be anti-microbial. Alternatively, an additional antimicrobial substance to be delivered to the microbial cells (e.g., an antibiotic) is included in the antimicrobial composition. By facilitating delivery to the target cells, the compounds can enhance the effectiveness of the additional antimicrobial substance. In some cases, the intensification can be substantial. Particularly important are target microbes (eg, gram-negative bacteria generally or bacteria having a substantial amount (> 40%) of a lipid A or lipid-like substance A in the outer membrane). Other microbes including fungi, viruses and yeasts can also be target organisms. The compounds may also be administered in other contexts to enhance cellular permeability to introduce any of a large number of different classes of substances into a cell, particularly the bacterial cells discussed above. In addition to introducing anti-microbial substances, the invention can be used to introduce other substances, such as macromolecules (eg, DNA without vector). In addition, compounds according to the invention can be used to permeabilize a sperm cell. The invention can also be used to make antimicrobial compositions (eg, disinfectants, antiseptics, antibiotics, etc.), which comprise one of the above compounds. These compositions are not limited to pharmaceuticals and can be used topically or in non-therapeutic contexts to control microbial (particularly bacterial) growth. For example, can be used in applications that kill or control microbes with the contact. In still another aspect, the invention generally characterizes methods for identifying compounds that are effective against a microbe by admiring a candidate compound and a compound according to the invention, and determining whether the candidate compound has a static or toxic effect (e.g. an antiseptic, germicidal, disinfectant or antibiotic effect) in the microbe. Again, bacteria such as those discussed above are preferred. This aspect of the invention permits useful tests of an extremely broad range of candidate antimicrobials, which are known to have an antimicrobial effect in some contexts, but which have not yet been shown to have any effect against certain classes of microbes, such like, the bacteria discussed before. As described in more detail below, this aspect of the invention allows testing a wide range of antibiotics that are currently thought to be ineffective against gram-negative bacteria or containing lipid-like substances A. In yet another aspect, the invention characterizes compositions that include one of the above compounds in combination with a substance to be introduced into a cell, such as an antimicrobial substance as described with more dellate above. The compound and the additional substance can be mixed with a pharmaceutically acceptable carrier. Other features or advantages of the present invention will be apparent from the following detailed description of various embodiments, and also from the appended claims. The invention encompasses steroid derivatives that can be made by the synthetic routes described herein, and methods for treating a subject having a condition mediated by a bacterial infection by administering an effective amount of a pharmaceutical composition containing a compound described herein to subject.

DESCRIPTION OF THE PREFERRED MODALITIES In general, the present invention provides the compounds of formula I described above. The methods of preparation and the M IC and MBC of compounds of formula I. The permeability of the cell membrane is also measured and described. Compounds that are useful according to the invention, as described below, include novel steroid derivatives that exhibit bacteriostatic, bactericidal and bacterial sensitizing properties. Those skilled in the art will appreciate that the invention extends to other compounds within the formulas given in the claims below, having the features described. These characteristics can be determined for each compound using the tests detailed below and elsewhere in the literature. Known compounds which are used according to the invention and precursors for novel compounds according to the invention can be purchased, for example, from Sigma Chemical Co., St. Louis; Aldrich, Milwaukee; Steroloíds and Research Plus. Other compounds according to the invention can be synthesized according to the known methods and the methods described below using publicly available precursors. The compounds of the present invention include, but are not limited to, amino or guanidino groups covalently bound to a steroid backbone, eg, cholic acid. The skeleton can be used to orient the amino or guanidino groups on one side of the steroid. Other steroid skeletons can also be used, for example, fused 5-membered rings. The biological activity of the compounds can be determined by standard methods known to those of skill in the art, such as, the "minimum inhibitory concentration (MIC)" assay described in the present examples, whereby MI C is recorded as lowest concentration at which no change in optical density (OD) is observed during a given period. When the compound is only tested against a control that lacks the compound, the antimicrobial effect of the compound is only determined. Alternatively, the "fractionated inhibitory concentration (FIC)" is also useful for the determination of synergy between the compounds of the invention, or the compounds in combination with known antibiotics. The FICs can be realized by means of board titrations of compounds in one dimension of a microtitre plate, and of antibiotics in the other dimension, for example. The FIC is calculated by looking at the impact of one antibiotic on the M IC of the other and vice versa. An ICF of one indicates that the influence of the compounds is additive and an ICF of less than one indicates synergy. Preferably, an FIC of less than 0.5 is obtained by synergism. As used herein, FIC may be determined as follows: FIC = M IC (compound in combination) + M IC (antibiotic in combination) M IC (compound only) M IC (antibiotic only) This procedure allows the determination of synergistic effects of the compound with other compounds. For example, substances that generally may not be sufficiently effective against certain bacteria in safe dosages may be made more effective with the compound of the invention, thereby allowing the use of the substances against new infection categories. Specifically, many existing antibiotics are effective against some gram-positive bacteria, but are not currently indicated to treat gram-negative bacterial infection. In some cases, the antibiotic may be ineffective by itself against gram negative bacteria because it fails to enter the cell. The compounds of the invention can increase the permeability in order to render the antibiotics effective against gram negative bacteria. In addition, the fractionated inhibitory concentration is also useful for the determination of synergy between the compounds of the invention in combination with other compounds having unknown antibacterial activity or in combination with other compounds, for example, compounds that have been tested and that show antibacterial activity. For example, compounds of the invention can increase the permeability in order to make compounds lacking antibacterial activity, effective against bacteria. The FIC can also be used to test other types of previously unappreciated activity of substances that will be introduced into the cell by means of permeability enhancing compounds according to the invention. Although we do not wish to unite to a single specific theory, and such theory is not necessary to practice the invention, a mechanism of action is the interaction of lipid A of multiple portions (usually three), which under physiological conditions are positively charged, for example , guanidino or amino portions. The portions extend from the general plane of the rest of the molecule, thus minimizing certain aspects of the structure of polyminines. In this regard, the compounds of the invention will generally be useful in the manner in which the polymyxins are useful.

Moreover, with respect to systemic administration, those skilled in the art will recognize screens of appropriate toxicity that allow the selection of non-toxic compounds in dosages that enhance microbial permeability. As noted, the invention also involves topical as well as non-therapeutic applications (antiseptic, germicidal or disinfectant), in which the compounds are contacted with surfaces to be treated. The term "contact" refers, preferably, to exposing the bacteria to the compound so that the compound can inhibit, kill or lyse bacteria, bind endotoxins (LPS), or permeabilize gram-negative bacterial outer membranes effectively. The contact can be in vitro, for example, by adding the compound to a bacterial culture to test the susceptibility of the bacterium to the compound. The contact may be in vivo, for example, by administering the compound to a subject with a bacterial disorder, such as septic shock. "Inhibit" or "effective inhibitory amount" refers to the amount of compound that is required to cause a bacteriostatic or bactericidal effect. Examples of bacteria that can be inhibited include E. coli, P. aeruginosa, E. cloacae, S. typhimurium, M. tuberculosis and S. aureus. The method for inhibiting the growth of bacteria may also include the addition of antibiotics by synergistic or combination therapy. The appropriate antibiotic administered normally will depend on the susceptibility of the bacterium, such as, whether the bacterium is gram negative or gram positive, and will be readily discernible by one of skill in the art. Examples of particular classes of antibiotics to be tested by synergistic therapy with the compounds of the invention (as described above) include aminoglycosides (e.g., tobramycin), penicillins (e.g., piperacillin), cephalosporins (e.g., ceftazidime), fluoroquinolones. (e.g., ciprofloxacin), carbepenems (e.g., imipenema), tetracyclines and macrolides (e.g., erythromycin and clarithromycin). The method of inhibiting the growth of bacteria may also include the addition of antibiotics for combination or synergistic therapy. The appropriate antibiotic commonly administered will depend on the susceptibility of the bacterium, such as whether the bacterium is gram negative or gram positive, and will be readily discernible by one of skill in the art. In addition to the antibiotics listed above, normal antibiotics include aminoglycosides (amikacin, gentamicin, kanamycin, netilmicin, tobramine, streptomycin, azithromycin, clarithromycin, erythromycin, stolate / ethylsuccinate / gluceptate / lactobionate / erythromycin stearate), beta-lactams, such as, penicillins (for example, penicillin G, penicillin V, methicillin, nafcillin, oxacillin, cloxacillin, dicloxacillin, ampicillin, amoxicillin, ticarcillin, carbenicillin, mezlocillin, azlocillin, and piperacillin), or cephalosporins (for example, cephalothin, cefazolin, cefaclor, cefamandole, cefoxitin, cefuroxime, cefonicid, cefmetazole, cefotetan, cefprozil, loracarbef, cefetamet, cefoperazone, cefotaxime, ceftizoxime, ceftriaxone, ceftazidime, cefepime, cefixime, cefpodoxime and cefsulodine). Other classes of antibiotics include carbapenems (e.g., imipenema), monbactams (e.g. aztreonam), quinolones (e.g., fleroxacin, nalidixic acid, norfloxacin, ciprofloxacin, ofloxacin, enoxacin, lomefloxacin, and cinoxacin), tetracyclines (e.g., doxycycline) , minocycline, tetracycline) and glycopeptides (for example, vancom icine, teicoplanin), for example. Other antibiotics include chloramphenicol, clindamycin, trimethoprim, sulfamethoxazole, nitrofurantoin, rifam pin, and mupirocin.

Administration The compounds can be administered to any host, including a human or non-human animal, in an amount effective to inhibit not only growth of a bacterium., but also a virus or fungus. These compounds are useful as antimicrobial agents, antiviral agents, spermicidal agents and antifungal agents. The compounds can be administered to any host, including a human or non-human animal, in an amount effective to inhibit not only growth of a bacterium, but also a virus or fungus. These compounds are useful as antimicrobial agents, antiviral agents and antifungal agents. The compounds of the invention can be administered parenterally by injection or by gradual infusion over time. The compounds can be administered topically, intravenously, intraperitoneally, intramuscularly, subcutaneously, intracavity or transdermally. Preferred methods for delivery of the compound include orally, by encapsulation in microspheres or proteinoids, by delivery of aerosol to the lungs, or transdermally by iontophoresis or transdermal electroporation. Other methods of administration will be known to those skilled in the art. Preparations for parenteral administration of a compound of the invention include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils, such as, olive oil, injectable organic esters, such as ethyl oleate. Aqueous carriers include water, solutions, emulsions or alcoholic / aqueous suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, fixed oils or lactated Ringer. Intravenous vehicles include nutrient and fluid fillers, electrolyte fillers (such as those based on Ringer's dextrose), and the like. The preservatives and other additives may also be present, such as, for example, antimicrobials, antioxidants, chelating agents and inert gases and the like. The invention provides a method for treating or ameliorating an endotoxemia or disorder associated with septic shock (sepsis), or one or more of the symptoms of sepsis comprising administering to a subject exhibiting the symptoms of sepsis or at risk of developing sepsis, an amount Therapeutically effective of a compound of the invention. The term "improve" refers to a decrease or decrease in the symptoms of the disorder being treated. Such symptoms, which may be improved include those associated with a transient increase in the blood level of TNF, such as fever, hypotension, neurotropenia, leukopenia, thrombocytopenia, disseminated intravascular coagulation, adult respiratory distress syndrome, shock or multiple failure of organs. Patients requiring such treatment include those at risk of or those suffering from, toxem ia, such as, endotoxemia resulting from an infection of gram-negative bacteria, poisoning or liver failure, for example. In addition, patients who have gram-positive, viral or fungal bacterial infection may exhibit symptoms of sepsis and may benefit from such a therapeutic method, as described herein. Those patients who are more particularly capable of benefiting from the method of the invention are those who suffer from infection by E. coli, Haemophilus influenza B, Neisseria meningitidis, staphylococci or pneumococci. Patients at risk for sepsis include those suffering from gunshot wounds, kidney or liver failure, trauma, burns, immunocompromised (HIV), hematopoietic neoplasms, multiple myeloma, Castleman's disease or cardiac myxoma. In addition, the compounds can be incorporated into biodegradable polymers that allow sustained release, the polymers being implanted in the vicinity of where delivery is desired, for example, at the site of a bacterial infection. The biodegradable polymers and their use are described in detail in Brem et al., J. Neurosurg, 74: 441-446 (1 991). As mentioned above, the present invention provides a pharmaceutical formulation having an effective amount of a compound of formula I for treating a patient having a bacterial infection. As used herein, an "effective amount" of the compound is defined as the amount of the compound upon which, upon administration to a patient, inhibits the growth of bacteria, kills bacterial cells, sensitizes bacteria to other antibiotics, or eliminates infection. bacterial in the treated patient. The dosage of the composition will depend on the condition being treated, the particular derivative used, and other clinical factors, such as, the weight and condition of the patient and the route of administration of the compound. However, for oral administration to humans, a dosage of 0.01 to 1000 mg / kg / day, preferably 0.01 -1 mg / kg / day, is generally sufficient. Effective doses will also vary, as will be recognized for those skilled in the art, depending on the route of administration, use of excipient, and the possibility of co-use with other therapeutic treatments including other antibiotic agents. For example, the term "therapeutically effective amount", as used herein for treatment of endotoxemia, refers to the amount of compound used which is sufficient to decrease the response of the subject to LPS and decrease the symptoms of sepsis. The term "therapeutically effective" therefore includes the amount of compound sufficient to prevent, and preferably reduce at least 50%, and more preferably sufficient to reduce by 90%, a clinically significant increase in the plasma level of TNF. . The dosage ranges for administration of the compound are those large enough to produce the desired effect. In general, the dosage will vary with the age, condition, sex and degree of infection with bacteria or other agents as described above, in the patient and can be determined by one skilled in the art. The dosage can be adjusted by the individual's physician in case of any contraindication. In any case, the effectiveness of treatment can be determined by monitoring the level of LPS and TNF in a patient. A decrease in serum LPS and TNF levels should correlate with patient recovery. In addition, patients at risk of or exhibiting the symptoms of sepsis can be treated by the method as described above, further comprising administering, substantially simultaneously with the therapeutic administration of the compound, a TNF inhibitor, an antibiotic or both. . For example, intervening in the role of TNF in sepsis, either directly or indirectly, such as, by the use of an anti-TNF antibody and / or a TNF antagonist, can prevent or ameliorate the symptoms of sepsis. Particularly preferred is the use of an anti-TNF antibody as an active ingredient, such as a monoclonal antibody with TNF specificity as described by Tracey, et al. (Nature, 330: 662, 1987). A patient who exhibits the symptoms of sepsis can be treated with an antibiotic in addition to the compound treatment. Normal antibiotics include an aminoglycoside, such as gentamicin or a beta-lactam, such as penicillin, or cephalosporin or any of the antibiotics as previously listed. Accordingly, a preferred therapeutic method of the invention includes administering a therapeutically effective amount of a cationic compound substantially simultaneously with the administration of a bactericidal amount of an antibiotic. Preferably, compound administration occurs within about 48 hours and preferably within about 2-8 hours, and most preferably, substantially concurrently with the administration of the anibiotic. The term "bactericidal amount" as used herein, refers to an amount sufficient to achieve a concentration of blood killing bacteria in the patient receiving the treatment. The bactericidal amount of antibiotic generally recognized as safe for administration to a human is well known in the art, and as is known in the art, varies with the specific antibiotic and the type of bacterial infection being treated. Due to the antibiotic, antimicrobial and antiviral properties of the compounds, they can also be used as preservatives or sterilizers of materials susceptible to microbial or viral contamination. The compounds of the invention can be used as broad spectrum antimicrobial agents directed to various specific applications. Such applications include the use of the compounds as preservatives in processed foods when they are verified as effective against organisms including Salmonella, Yersinia, Shigella, either alone or in combination with antibacterial food additives, such as, lysozymes.; as a topical agent (Pseudomonas, Streptococcus) and to kill odor-producing microbes (Micrococci). The relative effectiveness of the compounds of the invention for the described applications can be readily determined by one of skill in the art in determining the sensitivity of any organism to one of the compounds. Although focused primarily on classical gram-negative staining bacteria, whose outer capsule contains a substantial amount of lipid A, they can also be effective against other organisms with a hydrophobic outer capsule. For example, mycobacterium spp. have a protective waxy outer coating, and the compounds of the invention in combination with antibiotics, can provide enhanced effectiveness against Mycobacterial infection, including tuberculosis. In that case, the compounds could be administered by the nose (aspiration), by any of several known techniques. Apart from the antimicrobial action, the permeability provided by the compounds can intensify the introduction of a wide variety of substances into microbes. For example, the compounds can be used to enhance the introduction of macromolecules, such as DNA or RNA in microorganisms, particularly gram-negative bacteria. In that case, there may be no need for traditional vectors (eg, phages), used to pack nucleic acids when transfecting microbes. The conditions and techniques for introducing such macromolecules into microbes using the compounds of the invention will be routine in most cases. Formulations include those suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intraocular, intratracheal or epidural) administration. The formulations can be conveniently presented in unit dosage form and can be prepared by conventional pharmaceutical techniques. Such techniques include the step of bringing the active ingredient and the pharmaceutical carrier (s) into association. In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. Formulations of the present invention suitable for oral administration may be presented as discrete units, such as capsules or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil emulsion and as a bolus, etc. A tablet can be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing, in a suitable machine, the active ingredient in a free flowing form, such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface active agent or dispersant. The molded tablets can be made by molding, in a suitable machine, a mixture of the wetted powder compound with an inert liquid diluent. The tablets may optionally be coated or marked with lines, and may be formulated in order to provide a slow or controlled release of the active ingredient therein. Formulations suitable for topical administration in the mouth include tablets comprising the ingredients in a flavored base, usually sucrose and acacia or tragacanth; pills comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia; and mouth rinses comprising the ingredient to be administered in a suitable liquid carrier. Formulations suitable for topical administration to the skin can be presented as ointments, creams, gels and pastes comprising the ingredient to be administered in an acceptable pharmaceutical carrier. A preferred topical delivery system is a transdermal patch containing the ingredient to be administered. Formulations for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate. Formulations suitable for nasal administration, wherein the carriers are a solid, include a coarse powder having a particle size, for example, in the range of 20 to 500 microns, which is administered in the manner in which it is taken a sniff, that is, by rapid inhalation through the nasal passage from a powder container held near the nose. Suitable formulations, wherein the carrier is a liquid, for administration, such as, for example, a nasal spray, aerosol or nasal drops, include aqueous or oily solutions of the active ingredient.

Formulations suitable for vaginal administration can be presented as pessaries, tampons, creams, gels, pastes, foams or aerosols formulations containing, in addition to the active ingredient, carriers such as those known in the art to be appropriate. Formulations suitable for parenteral administration include sterile aqueous and non-aqueous injection solutions, which may contain anti-oxidants, buffers, other bacteriostats and solutes, which render the formulation isotonic with the intended recipient's blood; and sterile aqueous and non-aqueous suspensions, which may include suspending agents and thickening agents. The formulations can be presented in unit dose or multi-dose containers, for example, sealed vials and flasks, and can be stored under freeze-dried (lyophilized) conditions, which require only the addition of the sterile liquid carrier, eg, water for injections, immediately before use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit dosage formulations are those containing a daily dose or unit, daily sub-dose, as stated hereinbefore, or an appropriate fraction thereof, of the administered ingredient. It should be understood that, in addition to the ingredients, particularly mentioned above, the formulations of this invention may include other agents conventional in the art having regard to the type of formulation in question, for example, those suitable for oral administration, may include saborizatnes. The carrier in the pharmaceutical composition must be "acceptable" in the sense of being compatible with the active ingredient of the formulation (and preferably, capable of stabilizing it) and not be detrimental to the subject to be treated. Without further elaboration, it is believed that the foregoing description has adequately enabled the present invention. The following specific modalities are, accordingly, to be interpreted as merely illustrative, and not limiting of the rest of the description in any way. All publications cited herein, including patents, are incorporated herein by reference. Examples 1-6 represent the normal syntheses of compounds 1 to 48, as exemplified in Scheme 1 to 7. Example 7 represents the MIC and MCB tests. Example 8 represents the ability of the compounds of formula I to decrease the M ICs of other antibiotics. Example 9 represents other compounds of formula I, which can be synthesized using known starting materials and reaction schemes that are similar to those described herein. For example, the hydroxyl groups in cholic acid can be converted to amine by the method found in Hsieh et al. , Synthesis and DNA Binding Properties of C3-, C 1 2-, and C24- Substituted Amino-Steroids Derived from Bile Acids (Synthesis and DNA binding properties of amino substituted steroids of C3, C12 and C24, bile acid derivatives) , Biorganic and Medicinal Chemistry, 1995, vol.6, 823-838.

Scheme 1. Preparation of compounds 1, 2, 4 and 5.

Reagents (reaction yields in parentheses): a) LiAIH, THF (98%). b) trityl chloride, Et3N, DMF (70%). c) allyl bromide, NaH, THF (96%). d) O3, CH2Cl2, MeOH; Me2S; NaBH 4 (95%). e) 9-BBN, THF; H2O2, NaOH (80%). f) MsCI, CH2Cl2, Et3N (78%, 82%). g) NaN3, DMSO (66% for 20, 19 led directly to 23). h) TsOH, MeOH (94%, 94% overall of 19), i) MsCl, CH2Cl2, Et3N (99%, 97%). j) N-benzylmethylamine (95%, 96%). k) LiAIH4, THF (95%, 99%). I) NH2C (NH) SO3H, MeOH (91%, 89%).

Scheme 2. Preparation of compound 3.

Reagents (reaction yields in parentheses): a) KCN, DMSO; MeOH, TsOH (92%). b) MsCl, Et 3 N, CH 2 Cl 2; BnMeNH (88%). c) LiAIH4, AICI3, THF (50%).

Scheme 3. Preparation of 6 and 7.

Reagents (reaction yields in parentheses): a) dicyclohexylcarbodiimide, N-hydroxysuccinimide, methylphenylamine, CH2Cl2, MeOH (85%). b) LiAIH4, THF (82%). c) dicyclohexylcarbodiimide, dimethylaminopyridine, Boc-glycine, CH2Cl2 (68%). d) dicyclohexylcarbodiimide, dimethylaminopyridine, Boc-β-alanine, CH 2 Cl 2 (72%). e) HCl, dioxane (-100%, -100%).

Scheme 4. Synthesis of compound 8.

Reagents (reaction yields in parentheses): a) DIAD, Ph3P, p-nitrobenzoic acid, THF (85%); NaOH, MeOH (85%). b) allyl bromide, NaH, THF (79%). c) O3, CH2Cl2, MeOH; Me2S; NaBH 4 (65%). d) MsCl, CH2Cl2, Et3N (86%). e) NaN3, DMSO (80%). f) TsOH, MeOH (94%). g) MsCI, CH2Cl2, Et3N; N-benzylmethylamine (93%). g) LiAIH4, THF (94%).

Scheme 5. Synthesis of compounds 9 and 10.

Reagents (reaction yields in parentheses): a) NaH, octyl bromide, DMF (80%); LiAIH4, THF (60%). b) LiAIH4, THF (60%).

Scheme 6. Synthesis of compound 11.

Reagents (reaction yields in parentheses): a) ethylene glycol, p-toluenesulfonic acid, benzene; NaOH, MeOH (96%). b) allyl bromide, NaH, THF (90%). c) 9-BBN, THF; NaOH, H2O2, (54%). d) pyridinium p-toluenesulfonate, MeOH (98%). e) methanesulfonyl chloride, Et 3 N, CH 2 Cl 2; NaN3, MDSO (88%). f) LiAIH4, THF (69%).

Scheme 7. Synthesis of compound 12.

Reagents (reaction yields in parentheses): a) methanesulfonyl chloride, Et3N, CH2Cl2; NaBr, DMF (97%). b) 23, NaH, DMF (52%). c) LiAIH4, THF (76%).

Synthesis of Compounds 1-48 General: The 1H and 13C NMR spectra were recorded on a Varian Gemini 2000 (200 MHZ) or Varian Unity 300 (300 MHZ) spectrometer and are referenced to CHCI3 (1H) or CDCI3 (13C). The IR spectra were recorded on a Perkin Elmer 1600 FTIR instrument. The mass spectrometric data were obtained on a JOEL SX 102A spectrometer. THF was dried over Na / benzophenone and CH2Cl2 was dried over CaH2 before being used. The other reagents and solvents were obtained commercially and were used as received.

Example 1 Compound 13: Methyl cholate (30.67 g, 72.7 mmol) in dry THF (600 ml) and LiAIH4 (4.13 g, 109 mmol) were added to a 1 l round bottom flask. After refluxing for 48 hours, saturated aqueous Na 2 SO 4 (100 ml) was slowly introduced, and the resulting precipitate was filtered and waswith hot THF and MeOH. Recrystallization from MeOH gave colorless crystals of 13 (28.0 g, 98% yield). P.f. 236.5-238 ° C; IR (KBr) 3375, 2934, 1373, 1081 crtV1; 1 H NMR (CDCl 3 / MeOH-d 4, 200 MHZ) d 3.98 (bs, 1 H), 3.83 (bs, 1 H), 3.60-3.46 (m, 2 H), 3.38 (bs, 5 H), 2.30-2.10 (m, 2H), 2.05-1.05 (series of multiplets, 22 H), 1.03 (bs, 3 H), 0.92 (s, 3 H), 0.71 (s, 3 H); 13C NMR (CDCl3 / MeOH-d4, 50 MHz) d 73.89, 72.44, 68.99, 63.51, 48.05, 47.12, 42.49, 40.37, 39.99, 36.62, 36.12, 35.58, 35.40, 32.77, 30.69, 30.04, 29.02, 28.43, 27.27 , 23.96, 23.080, 18.00, 13.02; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 417.2992 (55.3%); cale.417.2981.

Compound 14: To a round bottom flask was added 13 (28.2 g, 71.7 mmol) in DMF (300 ml), Et3N (20 ml, 143.4 mmol), trityl chloride (25.98 g, 93.2 mmol) and DMAP (0.13 g). 1.07 mmol). The mixture was stirred at 50 ° C under N2 for 30 hours followed by the introduction of water (1000 ml) and extraction with EtOAc (5 x 200 ml). The combined extracts were waswith water and brine and then dried over MgSO. After solvent removal in vacuo, the residue was purified using SiO2 chromatography (CH2Cl2, Et2O and MeOH as levigantes) to give 14 as a pale yellow solid (31.9 g, 70% yield). P.f. 187 ° C (decomposition); IR (KBr) 3405, 2935, 1448, 1075 cm "1; 1 H NMR (CDCl 3, 200 MHz) d 7.46-7.42 (m, 6 H), 7.32-7.17 (m, 9 H), 3.97 (bs, 1 H ), 3.83 (bs, 1 H), 3.50-3.38 (m, 1H), 3.01 (bs, 1 H), 2.94 (dd, J = 14.2, 12.2 Hz, 2 H), 2.64 (bs, 1 H), 2.51 (bs, 1 H), 2.36-2.10 (m, 2 H), 2.00-1.05 (series of multiplets, 22 H), 0.96 (d, J = 5.8 Hz, 3 H), 0.87 (s, 3 H) 0.64 (s, 3 H); 13C NMR (CDCI3, 50MHZ) d 144.77, 128.93, 127.91, 127.01, 86.43, 73.35, 72.06, 68.66, 64.28, 47.47, 46.53, 41.74, 41.62, 39.64, 35.57, 35.46, 34.91, 34.82, 32.40, 30.55, 28.21, 27.69, 26.80, 26.45, 23.36, 22.59, 17.83, 12.61; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 659.4069 (100%); cale.659.4076.

Compound 15. To a round bottom flask was added 14 (20.0 g, 31.4 mmol) in dry THF (600 ml) and NaH (60% in mineral oil, 6.3 g, 157.2 mmol). The mixture was refluxed for 30 minutes under N2 followed by the addition of allyl bromide (27 ml, 314 mmol). After 60 hours under reflux, additional NaH (3 eq.) And allyl bromide (4 eq.) Were added. Following another 50 hours of reflux, water (20 ml) was introduced slowly followed by the addition of 1% HCl until the aqueous layer became neutral. The mixture was then extracted with ether (3 x 100 ml) and the combined extracts were washed with water (100 ml) and brine (2 x 100 ml). The ether solution was dried over anhydrous Na2SO4, and after solvent removal, the residue was purified using SiO2 chromatography (hexanes and EtOAc / hexanes 1: 8 as levigants) to give 15 (22.76 g, 96% yield) like a pale yellow crystal. IR (pure) 2930, 1448, 1087 cnT1; 1 H NMR (CDCl 3, 200 MHz) d 7.48-7.30 (m, 6 H), 7.32-7.14 (m, 9 H), 6.04-5.80 (m, 3 H), 5.36-5.04 (series of multiplets, 6 H), 4.14 -3.94 (m, 4 H), 3.74 (td, J = 13.8, 5.8 Hz, 2 H), 3.53 (bs, 1 H), 3.20-2.94 (m, 3 H), 3.31 (bs, 1 H), 2.38-1.90 (m, 4 H), 1.90-0.96 (series of multiplets, 20 H), 0.90 (d, J = 5.4 Hz, 3 H), 0.89 (s, 3 H), 0.64 (s, 3 H); 13C NMR (CDCI3, 50MHZ) d 144.83, 136.27, 136.08, 128.94, 127.90, 126.98, 116.46, 115.70, 86.42, 80.94, 79.29, 74.98, 69.52, 69.39, 68.86, 64.39, 46.51, 46.42, 42.67, 42.14, 39.92 , 35.63, 35.51, 35.13, 32.45, 28.98, 28.09, 27.66, 27.57, 26.72, 23.32, 23.11, 17.92, 12.69; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 779.5013 (86.1%); cale.779.5015.

Compound 16: To a three-necked round bottom flask was added 15 (3.34 g, 4. 4 mmol) in CH2Cl2 (200 ml) and methanol (100 ml). Through the cold solution (-78 ° C) ozone was bubbled through until a blue color persisted. The excess ozone was removed with oxygen flow. The mixture was left in a dry ice-acetone bath for one hour. Methyl sulfide (2.4 ml) was added and 15 minutes later, the mixture was treated with NaBH (1.21 g, 32 mmol) in 5% aqueous NaOH solution (10 ml) / methanol (10 ml) and allowed to warm room temperature. The mixture was washed with brine (3 x 50 ml) and the combined brine wash was extracted with CH2Cl2 (2 x 50 ml). The organic solution was dried over MgSO4. After chromatography of SiO2 (MeOH (5%) in CH2Cl2), 3.30 g (95% yield) of 16 was isolated as an oil. IR (pure) 3358, 2934, 1448, 1070 crn "1; 1H NMR (CDCI3, 200 MHZ) d 7.50-7.42 (m, 6 H), 7.32-7.17 (m, 9 H), 3.80-3.96 (series of multiplets, 20 H), 2.25-0.96 (series of multiplets, 24 H), 0.89 (bs, 6 H), 0.65 (s, 3 H); 13CNMR (CDCI3, 50 MHZ) d 144.73, 128.88, 127.87, 126.96, 86.38, 81.05, 79.75, 76.59, 70.33, 69.66, 69.30, 64.20, 62.25, 62.16, 62.03, 46.77, 46.36, 42.63, 41.77, 39.60, 35.43, 35.23, 35.05, 34.89, 32.42, 28.91, 27.93, 27.56, 27.15, 26.68, 23.35, 22.98, 22.85, 18.15, 12.60; HRFAB-MS (thioglycerol + Na + matrix) m / e: [(M + Na] +) 791.4860 (100%), cale.791.4863.

Compound 17: To a round bottom flask was added 16 (1.17 g, 1.55 mol) in dry THF (30 ml) under N 2 in an ice bath, followed by 9-BBN / THF solution (0.5 M, 10.2 ml, 5.51). mmol). The mixture was stirred at room temperature for 12 hours. Aqueous NaOH (20%) (2 ml) and hydrogen peroxide (30%) (2 ml) were added in sequence. The mixture was refluxed for 1 hour followed by the addition of brine (60 ml) and extraction with EtOAc (4 x 30 ml). The combined extracts were dried over anhydrous Na2SO4. The product (1.01 g, 80% yield) was obtained as a colorless oil after SiO2 chromatography (5% MeOH in CH2Cl2). IR (pure) 3396, 2936, 1448, 1365, 1089 crtV1; 1 H NMR (CDCl 3, 200 MHZ) d 7.50-7.42 (m, 6 H), 7.34-7.16 (m, 9 H), 3.90-3.56 (m, 13 H), 3.50 (bs, 1 H), 3.40-2.96 (series of multiplets, 6 H), 2.30-0.94 (series of multiplets, 30 H), 0.90 (s, 3 H), 0.88 (d, J = 5.4 Hz, 3 H), 0.64 (s, 3 H); 13C NMR (CDCI3, 50MHZ) d144.73, 128.88, 127.85, 126.94, 86.36, 80.52, 78.90, 76.36, 66.82, 66.18, 65.77, 64.22, 61.53, 61.41, 61.34, 46.89, 46.04, 42.60, 41.59, 39.60, 35.37 , 35.27, 34.88, 32.75, 32.44, 32.31, 28.82, 27.65, 27.48, 27.13, 26.77, 23.35, 22.74, 22.38, 18.08, 12.48; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 833.5331 (100%), cale. 833.5332.

Compound 18: To a round bottom flask was added 16 (3.30 g, 4.29 mmol) in CH2Cl2 (150 mL) and NEt3 (2.09 mL, 15.01 mmol). The mixture was placed in an ice bath under N2 followed by the addition of mesyl chloride (1.10 ml, 14.16 mmol). After 30 minutes, water (30 ml) and brine (200 ml) were added. The CH2Cl2 layer was washed with brine (2 x 50 ml) and dried over anhydrous Na2SO. The combined aqueous mixture was extracted with EtOAc (3 x 100 mL). The combined extracts were washed with brine and dried over anhydrous Na2SO4. The desired product (3.35 g, 78% yield) was isolated as a pale yellow oil after chromatography of SiO2 (EtOAc / hexanes 1: 1). IR (pure) 2937, 1448, 1352, 1174, 1120, 924 cm "1; 1 H NMR (CDCl 3, 200 MHZ) d 7.52-7.40 (m, 6 H), 7.34-7.20, (m, 9 H), 4.42 -4.24 (m, 6 H), 3.90-3.64 (m, 4 H), 3.60-3.30 (m, 4 H), 3.24-3.00 (m, 3 H), 3.10 (s, 6 H), 3.05 (s) , 3 H), 2.20-1.96 (m, 3 H), 0.91 (bs, 6 H, 0.65 (s, 3 H); 13 C NMR (CDCl 3, 50 MHz) d 114.68, 128.85, 127.85, 126.96, 86.37, 81.37 , 79.58, 76.58, 69.95, 69.43, 69.34, 66.52, 66.31, 65.59, 64.11, 46.80, 46.20, 42.65, 41.48, 39.35, 37.85, 37.48, 35.36, 34.92, 34.73, 32.37, 28.66.28.01, 27.44, 27.03, 26.72 , 23.17, 22.91, 22.72, 18.13, 12.50; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na) +] 1205.4176 (81.5%), step 1205.4189.

Compound 19: To a round bottom flask was added 17 (1.01 g, 1.25 mmol) in CH2CI2 (50 ml) and NEt3 (0.608 ml, 4.36 mmol). The mixture was placed in an ice bath under N2 followed by the addition of mesyl chloride (0.318 ml, 4.11 mmol). After 30 minutes, water (10 ml) and then brine (80 ml) were added. The CH2Cl2 layer was washed with brine (2 x 20 ml) and dried over anhydrous Na2SO. The combined aqueous mixture was extracted with EtOAc (3 x 40 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO4. The desired product (1.07 g, 82%) was isolated as a pale yellowish oil after SiO 2 chromatography (EtOAc / hexanes 1: 1). IR (pure) 2938, 1356, 1176, 1112 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.46-7.43, (m, 6 H), 7.32-7.22 (m, 9 H), 4.40-4.31 (m , 6H), 3.72-3.64 (m, 2 H), 3.55 (dd, J = 6.3, 5.8 Hz, 2 H), 3.51 (bs, 1 H), 3.32-3.14 (m, 3 H), 3.14-2.92 (m, 3 H), 3.01 (s, 3 H), 3.01 (s, 3 H), 3.00 (s, 3 H), 2.10-1.92 (m, 10 H), 1.92-1.58 (m, 8 H) , 1.56-0.92 (series of multiplets, 12 H), 0.90 (s, 3 H), 0.89 (d, J = 5.4 Hz, 3 H), 0.64 (s, 3 H); 13C NMR (CDCI3, 75 MHZ) d 144.67, 128.85, 127.85, 126.96, 86.42, 81.06, 79.83, 76.81, 68.12, 68.06, 68.02, 64.26, 36.06, 63.42, 46.76, 46.38, 42.73, 41.87, 39.73, 37.44, 37.32, 37.29, 35.52, 35.48, 35.32 , 35.06, 32.53, 30.55, 30.28, 30.02, 29.15, 27.96, 27.69, 27.61, 26.75, 23.52, 23.02, 18.17, 12.64; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 1067.4672 (100%), cale 1067.4659.

Compound 20: To a round bottom flask was added 18 (1.50 g, 1.50 mmol) in dry DMSO (20 ml) and NaN3 (0.976 g, 15 mmol). The mixture was heated to 80 ° C and stirred under N2 overnight, and then diluted with water (100 ml). The resulting aqueous mixture was extracted with EtOAc (3 x 50 mL), and the combined extracts were washed with brine and dried over anhydrous Na2SO. The desired product (0.83 g, 66% yield) was isolated as a clear crystal after chromatography of SiO2 (EtOAc / hexanes 1: 5). IR (pure) 2935, 2106, 1448, 1302, 1114 cm "1; 1 H NMR (CDCl 3, 200 MHz) d 7.50-7.42 (m, 6H), 7.36-7.20 (m, 9H), 3.84-3.70 (m, 2H), 3.65 (t, J = 4.9 Hz, 2H), 3.55 (bs, 1H), 3.44-3.08 (m, 10H), 3.02 (t, J = 6.4 Hz, 2H), 2.38-0.96 (series of multiplets , 24H), 0.92 (d, J = 5.6 Hz, 3H), 0.91 (s, 3H), 0.65 (s, 3H), 13C NMR (CDCI3, 50MHZ) d 114.84, 128.97, 127.92, 126.99, 86.42, 81.24 , 80.12, 76.59, 67.84, 67.29, 66.66, 64.36, 51.67, 51.44, 51.18, 46.53, 46.23, 42.21, 41.93, 39.73, 35.66, 35.36, 35.06, 34.78, 32.40, 28.95, 27.76, 27.39, 26.87, 23.45, 22.98 22.92, 17.98, 12.53; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + Na] +) 866.5040 (100%), cale.866.5057.

Compound 22: To a round bottom flask were added 20 (830 mg, 0.984 mmol) in MeOH (30 ml) and CH2Cl2 (30 ml) and p-toluenesulfonic acid (9.35 mg, 0.0492 mmol). The solution was stirred at room temperature for 2.5 hours, and then saturated aqueous NaHCO3 (10 ml) was introduced. Brine (30 ml) was added, and the mixture was extracted with EtOAc (4 x 20 ml). The combined extracts were dried over anhydrous Na2SO. The desired product (0.564 g, 95% yield) was isolated as a pale yellowish oil after chromatography of SiO2 (EtOAc / hexanes 1: 2). IR (pure) 3410, 2934, 2106, 1301, 1112, cm "1; 1 H NMR (CDCl 3, 200 MHz) d 3.80-3.54 (m, 7 H), 3.44-3.20 (m, 10 H), 2.35-0.96 (series of multiplets, 24 H), 0.95 (d, J = 6.4 Hz, 3 H) , 0.92 (s, 3H), 0.68 (s, 3H); 13C NMR (CDCI3, 50MHZ) d 81.10, 80.01, 76.60, 67.75, 67.16, 66.56, 63.63, 51.57, 51.34, 51.06, 46.29, 46.12, 41.81, 39.60, 35.55, 35.23, 34.94, 34.66, 31.75, 29.48, 28.81 , 27.72, 27.66, 27.29, 23.32, 22.86, 22.80, 17.85, 12.39; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + Na] +) 624.3965 (100%), cale.624.3962 Compound 23: To a round bottom flask was added 19 (1.07 g, 1025 mmol) and NaN3 (0.666 g, 10.25 mmol) followed by the introduction of dry DMSO (15 ml). The mixture was heated to 80 ° C under N2 overnight. After the addition of H2O (100 ml), the mixture was extracted with EtOAc (4 x 40 ml) and the combined extracts were washed with brine (2 x 50 ml) and dried over anhydrous Na2SO4. After solvent removal, the residue was dissolved in MeOH (15 ml) and CH2CI2 (15 ml) followed by the addition of catalytic amount of p-toluenesulfonic acid (9.75 mg, 0.051 mmol). The solution was stirred at room temperature for 2.5 hours before the addition of saturated NaHCO3 solution (15 ml). After the addition of brine (60 ml), the mixture was extracted with EtOAc (5 x 30 ml). The combined extracts were washed with brine (50 ml) and dried over anhydrous Na2SO4. The desired product (0.617 g, 94% yield over two steps) was obtained as a yellowish oil after SiO2 chromatography (EtOAc / hexanes 1: 2). IR (pure) 3426, 2928, 2094, 1456, 1263, 1107, crn "1; 1H NMR (CDCI3, 300 MHZ) d 3.68-3.56 (m, 3H, 3.56 - 3.34 (series of multiplets, 10H), 3.28- 3.00 (series of multiplets, 4H), 2.20-2.00 (m, 3H), 1.98-1.55 (series of multiplets, 15H), 1.55-0.96 (series of multiplets, 13H), 0.92 (d, J = 6.6 Hz, 3H ), 0.89 (s, 3H), 0.66 (s, 3H), 13C NMR (CDCI3, 75MHZ) d 80.63, 79.79, 76.04, 64.99, 64.45, 64.30, 63.72, 49.01, 48.94, 48.74, 46.49, 46.39, 42.70 , 41.98, 39.80, 35.65, 35.42, 35.28, 35.08, 31.99, 29.78, 29.75, 29.70, 29.49, 29.06, 27.87, 27.79, 27.65, 23.53, 23.04, 22.85, 18.05, 12.59; HRFAB-MS (thioglycerol + Na matrix) m / e: ([M + Na] +) 666.4415 (100%), cale.666.4431.

Compound 24: To a round bottom flask was added 22 (0.564 g, 0.938 mmol) in CH2Cl2 (30 ml) and NEt3 (0.20 ml, 1.40 mmol). The mixture was placed in an ice bath under N2 followed by the addition of mesyl chloride (0.087 ml, 1.13 mmol). After 30 minutes, water (20 ml) and brine (100 ml) were added. The CH2Cl2 layer was washed with brine (2 x 20 ml) and dried over anhydrous Na2SO4. The combined aqueous mixture was extracted with EtOAc (3 x 30 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO. The desired product (0.634 g, 99% yield) was isolated as a pale yellowish oil after chromatography of SiO2 (EtOAc / hexanes 1: 2). IR (pure) 2935, 2106, 1356, 1175, 1113, cm'1; H NMR (CDCI3, 300MHZ): d 4.20 (t, J = 6.8 Hz, 2H), 3.80-3.75 (m, 1H), 3.70-3.64 (m, 3H), 3.55 (bs, 1H), 3.44-3.01 ( m, 10H), 3.00 (s, 3H), 2.32-2.17 (m, 3H), 2.06-2.03 (m, 1H), 1.90-0.88 (series of multiplets, 20H), 0.95 (d, J = 6.6 Hz, 3H), 0.91 (s, 3H), 0.68 (s, 3H); 13C NMR (CDCI3, 75MHZ) d 80.90, 79.86, 76.43, 70.78, 67.64, 66.99, 66.48, 51.50, 51.26, 50.97, 46.05, 45.96, 42.08, 41.71, 39.51, 37.33, 35.15, 34.86, 34.60, 31.34, 28.73 , 27.62, 27.59, 27.51, 25.68, 23.22, 22.80, 22.70, 17.62, 12.33; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 702.37.41 (100%), cale.702.3737.

Compound 25: To a round bottom flask was added 23 (0.617 g, 0.96 mmol) in CH2Cl2 (30 ml) and NEt3 (0.20 ml, 1.44 mmol). The mixture was placed in an ice bath under N2 followed by the addition of mesyl chloride (0.089 ml, 1.15 mmol). After 30 minutes, water (20 ml) and brine (120 ml) were added. The CH2Cl2 layer was washed with brine (2 x 20 ml) and dried over anhydrous Na2SO4. The combined aqueous mixture was extracted with EtOAc (3 x 30 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO4. The desired product (0.676 g, 97% yield) was isolated as a pale yellowish oil after solvent removal. IR (pure) 2934, 2094, 1454, 1360, 1174, 1112 cm'1; 1 H NMR (CDCl 3, 300 MHZ) d 4.17 (t, J = 6.6 Hz, 2 H), 3.65-3.28 (series of multiplets, 11 H), 3.64-3.00 (series of multiplets, 4 H), 2.97 (s, 3 H) , 2.18-1.96 (series of multiplets, 16H), 1.54-0.94 (series of multiplets, 11H), 0.89 (d, J = 6.6 Hz, 3 H), 0.86 (s, 3H), 0.63 (s, 3H); 13C NMR (CDCI3, 75MHZ) d 80.47, 79.67, 75.92, 70.84, 64.90, 64.37, 64.17, 48.90.48.86, 48.66, 46.32, 46.26, 46.63, 41.87, 39.70, 37.39, 35.34, 35.28, 35.20, 34.99, 31.61 , 29.68, 29.60, 28.96, 27.78, 27.68, 27.57, 25.79, 23.41, 22.95, 22.74, 17.82, 12.50; HRFAB-MS (thioglycerol matrix) m / e: ([M + H] +) 722.4385 (22.1%), cale.722.4387.

Compound 26: To a 50 ml round bottom flask was added 24 (0.634 g, 0.936 mmol) and N-benzylmethylamine (2 ml). The mixture was heated under N2 at 80 ° C overnight. Excess N-benzylmethylamine was removed under vacuum, and the residue was subjected to SiO2 chromatography (EtOAc / hexanes 1: 2). The desired product (0.6236 g, 95% yield) was isolated as a pale yellow oil. IR (pure) 2935, 2106, 1452, 1302, 1116 cm "1; 1H NMR (cDCI3, 200 MHZ) d 7.32-7.24 (m, 5H), 3.80-3.76 (m, 1H), 3.70-3.60 (m, 3H), 3.54 (bs, 1H), 3.47 (s, 2H), 3.42-3.10 (m, 10 H), 2.38-2.05 (m, 5H), 2.17 (s, 3H), 2.02-0.88 (series of multiplets , 21 H), 0.93 (d, J = 7.0 Hz, 3 H), 0.91 (s, 3 H), 0.66 (s, 3 H); 3 C NMR (CDCl 3, 50 MHz) d 139.60, 129.34, 128.38, 127.02, 81.22, 80.10, 76.71, 67.85, 67.29, 66.65, 62.45, 58.38, 51.65, 51.44, 51.16, 46.50, 46.21, 42.40, 42.20, 41.93, 39.72, 35.80, 35.34, 35.05, 34.76, 33.65, 28.93, 27.82, 27.75, 27.38, 24.10, 23.45, 22.98, 22.91, 18.05, 12.50; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM-H] +) 703.4748 (90.2%), block 703.4772; QM + H] +) 705.4911 (100 %), cale.705.4928; ([M + Na] +) 727.4767 (1.5%), cale.727.4748.

Compound 27: To a 50 ml round bottom flask was added 25 (0.676 g, 0.937 mmol) and N-benzylmethylamine (2 ml). The mixture was heated under N2 at 80 ° C overnight. The excess of N-benzylmethylamine was removed under vacuum and the residue was subjected to SiO 2 chromatography (EtOAc / hexanes 1: 2). The desired product (0.672 g, 96% yield) was isolated as a pale yellow oil. IR (pure) 2934, 2096, 1452, 1283, 1107 cm "1; 1H NMR (CDCI3, 300 MHz) d 7.34-7.20 (m, 5H), 3.68-3.37 (series of multiplets, 13H), 3.28-3.04 ( m, 4H), 2.33 (t, J = 7.0 Hz, 2H), 2.18 (s, 3H), 2.20-2.00 (m, 3H), 1.96-1.56 (series of multiplets, 14 H), 1.54-1.12 (m , 10H), 1.10-0.96 (m, 3 H), 0.91 (d, J = 8.7 Hz, 3H), 0.89 (s, 3H), 0.65 (s, 3H); 13C NMR (CDCI3, 75 MHz) d 139.48 , 129.23, 128.30, 126.96, 80.66, 79.81, 7.08, 65.00, 64.46, 644.34, 62.50, 58.37, 49.02, 48.95, 48.75, 46.65, 46.40, 42.69, 42.43, 42.00, 39.83, 35.86, 35.45, 35.30, 35.10, 33.83 , 29.81, 29.78, 29.72, 29.09, 27.88, 27.81, 27.66, 24.19, 23.57, 23.06, 22.87, 18.15, 16.62; HRFAB-MS (thioglycerol matrix) m / e: ([M + H] +) 747.5406 (77.2% ), cale.747.5398.

Compound 1: To a round bottom flask was added 26 (0.684 g, 0.971 mmol) in dry THF (30 ml) and LiAIH4 (113.7 mg, 3.0 mmol) under N2. The mixture was stirred at room temperature for 13 hours, and then Na 2 SO 10 H 2 O powder (10 g) was added slowly. After the gray color disappeared, the mixture was filtered through Celite and washed with dry THF. The product (0.581 g, 95% yield) was obtained as a colorless crystal. IR (pure) 33.72, 2937, 1558, 1455, 1362, 1102 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.34-7.20 (m, 5H), 3.68-3.48 (m, 5H), 3.47 (s, 2H), 3.29 (bs, 1H), 3.22-3.00 (m, 3H), 2.96-2.80 (m, 6H), 2.32 (t, J = 6.8, 5.4 Hz, 2H), 2.17 (s, 3H), 2.20 -2.00 (m, 3H), 1.96-0.96 (series of multiplets, 27 H), 0.93 (d, J = 6.8 Hz, 3H), 0.90, (s, 3H), 0.67 (s, 3H), 13C NMR ( CDCI3, 75 MHz) d 139.50, 129.22, 128.31, 126.96, 80.76, 79.86, 76.10, 70.90, 70.33, 70.24, 62.48, 58.27, 46.55, 46.45, 42.72, 42.58, 42.33, 41.99, 39.77, 35.78, 35.37, 35.01, 33.73, 29.07, 27.95, 27.71, 24.06, 23.46, 22.99, 18.14, 12.55; HRFAB-MS (thioglycerol matrix) m / e: ([M + H] +) 627.5211 (100%), cale.627.5213.

HCl salt of compound 1: Compound 1 was dissolved in a minimum amount of CH 2 Cl 2 and excess HCl in ether was added. The solvent and excess HCl were removed in vacuo and a non-crystalline white powder was obtained. 1 H NMR (methanol-d 4/15% CDCl 3, 300 MHz) d 7.61-7.57 (m, 2 H), 7.50-7.48 (m, 3 H), 4.84 (bs, 10 H), 4.45 (bs, 1 H), 4.30 (bs, 1H), 3.96-3.82 (m, 2H), 3.78-3.69 (m, 2H), 3.66 (bs, 1H), 3.59-3.32 (series of multiplets, 4H), 13.28-3.02 (m, 8H) , 2.81 (s, 3H), 2.36-2.15 (, 4H), 2.02-1.68 (m, 8H), 1.64-0.90 (series of multiplets, 12 H), 1.01 (d, J = 6.35 Hz, 3 H), 0.96 (s, 3H), 0.73 (s, 3H); 13C NMR (methanol-d4 / 15% CDCI3, 75 MHz) d 132.31, 131.20, 130.92, 130.40, 83.13, 81.09, 78.48, 65.54, 64.98, 64.11, 60.87, 57.66, 47.51, 46.91, 43.52, 43.00, 41.38, 41.19, 41.16, 40.75, 40.30, 36.37, 36.08 , 36.00, 35.96, 33.77, 29.68, 29.34, 28.65, 28.37, 24.42, 24.25, 23.33, 21.51, 18.80, 13.04, Compound 2: To a round bottom flask were added 27 (0.82 g, 1.10 mmol) in dry THF ( 150 ml) and LiAIH4 (125 mg, 3.30 mmol) under N2. The mixture was stirred at room temperature for 12 hours and Na2SO-10 H2O powder (10 g) was slowly added. After the gray color disappeared, the mixture was filtered through a cotton plug and washed with dry THF. THF was removed in vacuo and the residue was dissolved in CH2Cl2 (50 mL). After filtration, the desired product was obtained as a colorless crystal (0.73 g, 99% yield). IR (pure) 3362, 2936, 2862, 2786, 1576, 1466, 1363, 1103 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.32-7.23 (m, 5 H), 3.67-3.63 (m, 1 H), 3.60-3.57 (m, 1 H), 3.53 (t, J = 6.4 Hz, 2 H), 3.47 ( s, 2H), 3.46 (bs, 1H), 3.24-3.17 (m, 2H), 3.12-2.99 (m, 2H), 2.83-2.74 (series of multiplets, 6H), 2.30 (t, J = 7.3 Hz, 2H), 2.15 (s, 3H), 2.20-2.00 (m, 3H), 1.95-1.51 (series of multiplets, 20 H), 1.51-1.08, (series of multiplets, 10 H), 1.06-0.80 (m, 3H), 0.87 (d, J = 8.1 Hz, 3H), 0.86 (s, 3H), 0.61 (s, 3H); 13C NMR (CDCI3, 75 MHz) d 139.35,129.16, 128.22, 126.88, 80.44, 79.29, 75.96, 66.70, 66.52, 66.12, 62.45, 58.26, 46.76, 46.27, 42.69, 42.41, 42.02, 40.68, 40.10, 40.02, 39.82 , 35.84, 35.47, 35.30, 35.06, 34.15, 34.09, 34.03, 33.80, 28.96, 27.93, 27.75, 27.71, 24.32, 23.53, 23.03, 22.75, 18.17, 12.58; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 691.5504 (38.5%), cale.691.5502.

HCl salt of compound 2: Compound 2 was dissolved in a minimum amount of CH 2 Cl 2 and excess HCl in ether was added. Removal of the solvent and excess HCl gave a non-crystalline white powder. 1H NMR (methanol-d4 / 15% CDCl 3, 300 MHz) d 7.60-7.59 (m, 2H), 7.50-7.47 (m, 3H), 4.82 (bs, 10H), 4.43 (bs, 1H), 4.32 (bs , 1H), 3.85-3.79 (m, 1H), 3.75-3.68 (m, 1H), 3.64 (t, J = 5.74 Hz, 2H), 3.57 (bs, 1H), 3.36-3.28 (m, 2H), 3.25-3.00 (multitude series, 10 H), 2.82 (s, 3H), 2.14-1.68 (series of multiplets, 19H), 1.65-1.15 (series of multiplets, 11 H), 0.98 (d, J = 6.6 Hz, 3H), 0.95 (s, 3H), 0.72 (s, 3H); 13C NMR (methanol-d4 / 15% CDCl3, 75 MHz) d 132.21, 131.10, 130.58, 130.28, 81.96, 80.72, 77.60, 66.84, 66.58, 66.12, 61.03, 57.60, 44.16, 42.77, 40.62, 39.57, 39.43, 36.28 , 36.03, 35.96, 35.78, 33.65, 29.48, 29.27, 29.11, 29.01, 28.61, 28.56, 28.35, 24.25, 23.56, 23.30, 21.17, 18.64, 12.90.

Compound 4: A suspension of 1 (79.1 mg, 0.126 mmol) and aminoiminomethanesulfonic acid (50.15 mg, 0.404 mmol) in methanol and chloroform was stirred at room temperature for 24 hours, and the suspension became clear. An ether solution of HCl (1 M, 1 ml) was added followed by the removal of solvent with N2 flow. The residue was dissolved in H2O (5 ml) followed by the addition of 20% aqueous NaOH (0.5 ml). The resulting nebula mixture was extracted with CH2Cl2 (4 x 5 ml). The combined extracts were dried over anhydrous Na2SO4. Removal of solvent gave the desired product (90 mg, 95%) as a white powder. P.f. 111-112 ° C. IR (pure) 3316, 2937, 1667, 1650, 1556, 1454, 1348, 1102 cm "1; 1 H NMR (5% methanol-d4 / CDCl 3, 300 MHz) d 7.26-7.22 (m, 5H), 4.37 (bs , 3H), 3.71-3.51 (series of multiplets, 5H), 3.44 (s, 2H), 3.39-3.10 (series of multiplets, 10 H), 2.27 (t, J = 6.83 Hz, 2H), 2.13 (s, 3H), 2.02-0.94 (series of multiplets, 33 H), 0.85 (d, J = 5.62 Hz, 3H), 0.84 (s, 3H), 0.61 (s, 3H); 13C NMR (5% methanol-d4 / CDCI3, 75 MHz) d 158.54, 158.48, 158.43, 138.27, 129.47, 128.32, 127.19, 81.89, 80.30, 77.34, 69.02, 68.46, 67.21, 62.36, 58.00, 47.36, 46.18, 43.26, 43.00, 42.73, 42.18, 41.48, 39.32, 35.55, 34.97, 34.89, 34.67, 33.63, 28.93, 28.28, 27.53, 27.16, 23.96, 23.28, 23.16, 22.77, 18.36, 12.58; HRFAB-S (thioglycerol + Na + matrix) m / e: QM + H] + ) 753.5858 (100%), cale.753.5867.

HCl salt of compound 4: Compound 4 was dissolved in minimal amount of CH 2 Cl 2 and MeOH followed by addition of excess HCl in ether. The solvent was removed by N2 flow, and the residue was subjected to high vacuum overnight. The desired product was obtained as a non-crystalline white powder. 1 H NMR (methanol-d 4/20% CDCl 3, 300 MHz) d 7.58 (bs, 2 H), 7.50-7.48 (, 3 H), 4.76 (bs, 13 H), 4.45 (d, J = 12.9 Hz, 1 H), 4.27 (dd, 1h, J = 12.9, 5.4 Hz), 3.82-3.00 (series of multiplets, 17 H), 2.81-2.80 (m, 3H), 2.20-1.02 (series of multiplets, 27 H), 0.98 (d) , J = 6.59 Hz, 3H), 0.95 (s, 3H), 0.72 (s, 3H); 13C NMR (methanol-d4 / 20% CDCl3, 75 MHz) d158.88, 158.72, 132.00, 131.96, 130.98, 130.15, 82.51, 81.07, 78.05, 68.50, 68.02, 67.94, 67.10, 60.87, 60.53, 57.38, 47.16, 46.91, 43.91, 43.11, 43.01, 42.91, 42.55, 40.28, 39.88, 39.95, 35.90, 35.73, 35.64, 33.53, 29.18, 28.35, 27.99, 24.02, 23.30, 21.35, 18.52, 18.44, 13.06.

Compound 5: A suspension of 2 (113 mg, 0.169 mmol) and aminoiminomethanesulfonic acid (67.1 mg, 0.541 mmol) in methanol and chloroform was stirred at room temperature for 24 hours. HCl in ether (1 M, 1 ml) was added followed by solvent removal with N2 flow. The residue was subjected to high vacuum overnight and dissolved in H2O (5 mL) followed by the addition of 20% NaOH solution (1.0 mL). The resulting mixture was extracted with CH2Cl2 (5 x 5 ml). The combined extracts were dried over anhydrous Na2SO. Removal of solvent gave the desired product (90 mg, 95% yield) as a white solid. Mp. 102-104 ° C. IR (pure) 3332, 3155, 2939, 2863, 1667, 1651, 1558, 1458, 1350, 1100 cm "1; 1 H NMR (5% methanol-d 4 / CDCl 3, 300 MHz) d 7.35-7.24 (m, 5H) , 3.75-3.64 (m, 1H), 3.57 (bs, 5H), 3.50 (s, 2H), 3.53-3.46 (m, 1H), 3.40-3.10 (series of multiplets, 14 H), 2.34 (t, J = 7.31 Hz, 2H), 2.19 (s, 3H), 2.13-0.96 (series of multiplets, 36 H), 0.91 (bs, 6H), 0.66 (s, 3H); 13C NMR (5% methanol-d4 / CDCl3, 75 MHz) d 157.49, 157.31, 157.23, 138.20, 129.52, 128.34, 127.23, 81.17, 79.19, 76.42, 65.63, 65.03, 64.70, 2.70, 62.36, 58.02, 47.23, 46.24 , 42.89, 42.18, 41.45, 39.45, 39.40, 39.30, 38.71, 35.61, 35.55, 35.02, 34.82, 33.69, 29.87, 29.59, 29.42, 28.84, 27.96, 27.56, 23.95, 23.40, 22.82, 22.64, 18.28, 12.54; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 795.6356 (84.3%), cale.795.6337.

HCl salt of compound 5: Compound 5 was dissolved in a minimum amount of CH 2 Cl 2 and MeOH followed by the addition of excess HCl in ether. The solvent and excess HCl were removed by N2 flow and the residue was subjected to high vacuum overnight. The desired product was obtained as a non-crystalline white powder. H NMR (methanol-d4 / 10% CDCl 3, 300 MHz) d 7.62-7.54 (m, 2H), 7.48-7.44 (m, 3H), 4.84 (bs, 16 H), 4.46 (d, J = 12.7 Hz, 1H), 4.26 (dd, J = 12.7, 3.42 Hz, 1H), 3.78-3.56 (series of multiplets, 5 H), 3.38-3.05 (series of multiplets, 13H), 2.80 (d, 3H), 2.19-2.04 (m, 3H), 2.02-1.04 (series of multiplets, 30 H), 0.98 (d, J = 6.35 Hz, 3H), 0.95 (s, 3H), 0.72 (s, 3H); 13 C NMR (methanol-d 4/10% CDCl 3, 75 MHz) d 158.75, 158.67, 132.32, 131.24, 130.83, 130.43, 82.49, 81.02, 77.60, 66.47, 65.93, 61.19, 60.85, 57.69, 47.79, 47.60, 44.29, 43.07 , 40.86, 40.42, 40.19, 40.09, 39.76, 36.68, 36.50, 36.15, 35.94, 33.91, 30.75, 30.46, 29.74, 29.33, 28.71, 24.41, 24.03, 23.38, 22.21, 22.16, 18.59, 18.52, 13.09.

Example 2 Compound 28: A suspension of 19 (0.641 g, 0.614 mmol) and KC (0.40 g, 6.14 mmol) in anhydrous DMSO (5 mL) was stirred under N2 at 80 ° C overnight, followed by the addition of H2O (50 ml). The aqueous mixture was extracted with EtOAc (4 x 20 ml). The combined extracts were washed with brine once, dried over anhydrous Na2SO and concentrated in vacuo. The residue was dissolved in CH2Cl2 (3 mL) and MeOH (3 mL) and a catalytic amount of p-toluenesulfonic acid (5.84 mg, 0.03 mmol) was added. The solution was stirred at room temperature for 3 hours before the introduction of saturated NaHCO3 solution (10 ml). After addition of brine (60 ml), the mixture was extracted with EtOAc (4 x 30 ml). The combined extracts were washed with brine once and dried over anhydrous Na2SO4 and concentrated. The residue gave the desired product (0.342 g, 92% yield) as pale yellowish oil after column chromatography (silica gel, EtOAc / hexanes 2: 1). IR (pure) 3479, 29.36, 2864, 2249, 1456, 1445, 1366, 1348, 1108 cm "1; 1H NMR (CDCI3, 300 MHz) d 3.76-3.53 (m, 7H), 3.32-3.06 (series of multiplets , 4H), 2.57-2.46 (m, 6 H), 2.13-1.00 (series of multiplets, 31 H), 0.93 (d, J = 6.35 Hz, 3H), 0.90 (s, 3H), 0.67 (s, 3H) ); 3C NMR (CDCI3, 75 MHz) d 119.91, 119.89, 80.75, 79.65, 76.29, 65.83, 65.37, 65.19, 63.63, 46.57, 46.44, 42.77, 41.79, 39.71, 35.63, 35.26, 35.02, 32.00, 29.46, 29.03 , 27.96, 27.74, 26.64, 26.42, 26.12, 23.56, 22.98, 22.95, 18.24, 14.65, 14.54, 14.30, 12.60; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 618.4247 ( 67.8%), cale.618.4247.

Compound 29: To a solution of 28 (0.34 g, 0.57 mmol) in dry CH 2 Cl 2 (15 ml) under N 2 at 0 ° C, NEt 3 (119.5 μl, 0.857 mmol) was added followed by the addition of mesyl chloride (53.1 μl). 0.686 mmol). The mixture was allowed to stir at 0 ° C for 30 minutes before the addition of H2O (6 ml). After the introduction of brine (60 ml), the aqueous mixture was extracted with EtOAc (4 x 20 ml). The combined extracts were washed with brine once, dried over anhydrous Na2SO and concentrated. To the residue was added N-benzylmethylamine (0.5 ml) and the mixture was stirred under N2 at 80 ° C overnight. The excess N-benzylmethylamine was removed in vacuo and the residue was subjected to column chromatography (silica gel, EtOAc / hexanes 2: 1 followed by EtOAc) to give the product (0.35 g, 88% yield) as an oil. pale yellow. IR (pure) 2940, 2863, 2785, 2249, 1469, 1453, 1366, 1348, 1108 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.34-7.21 (m, 5H), 3.76-3.69 (m, 1H ), 3.64-3.50 (m, 4H), 3.48 (s, 2H), 3.31-3.05 (series of multiplets, 4H), 2.52-2.46 (m, 6H), 2.33 (t, J = 7.32 H, 2Hz), 2.18 (s, 3H), 2.13-0.95 (series of multiplets, 30 H), 0.91 (d, J = 6.80 H, 3 Hz), 0.90 (s, 3H), 0.66 (s, 3H); 13C NMR (CDCI3 , 75 MHz) d 139.37, 129.17, 128.26, 126.93, 119.96, 119.91, 80.73, 79.59, 76.26, 65.79, 65.35, 65.13, 62.47, 58.25, 46.74, 46.40, 42.72, 42.38, 41.76, 39.68, 35.78, 35.22, 34.98 , 33.79, 28.99, 27.92, 27.71, 26.63, 26.38, 26.09, 24.21, 23.54, 22.96, 22.90, 18.28, 14.62, 14.51, 14.26, 12.58; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 699.5226 (100%), cale.699.5213.

Compound 3: A solution of 29 (0.074 g, 0.106 mmol) in anhydrous THF (10 mL) was added dropwise to a mixture of AICI3 (0.1414 g, 1.06 mmol) and LiAIH4 (0.041 g, 1.06 mmol) in THF dry (10 ml). The suspension was stirred for 24 hours followed by the addition of 20% aqueous NaOH solution (2 ml) at ice bath temperature. Anhydrous Na2SO4 was added to the aqueous paste. The solution was filtered and the precipitate was washed twice with THF. After solvent removal, the residue was subjected to column chromatography (silica gel, MeOH / CH 2 Cl 2 1: 1 followed by MeOH / CH 2 Cl 2 / NH 3 H 2 O 4: 4: 1) to give the desired product 0.038 g, 50% of performance) as a transparent oil. IR (pure) 3362, 2935, 2863, 2782, 1651, 1574, 1568, 1557, 1471, 1455, 1103 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.32-7.22 (m, 5H), 3.60-3.02 (series of wide multiplets, 18 H), 2.90-2.70 (m, 5H), 2.33 (t, J = 7.20 Hz, 2H), 2.24-2.04 (m, 3H), 2.18 (s, 3H), 1.96-0.96 (series of multiplets, 30 H), 0.90 (d, J = 7.57 Hz, 3H), 0.89 (s, 3H), 0.64 (s, 3H); 13C NMR (CDCI3, 75 MHz) d 139.44, 129.24, 128.31, 126.97, 80.63, 79.65, 75.97, 68.44, 68.00, 67.96, 62.54, 58.40, 46.77, 46.30, 42.73, 42.43, 42.07, 41.92, 41.74, 41.72, 39.81, 35.82, 35.48, 35.07, 33.84, 31.04, 30.30, 30.10, 29.03, 28.11, 27.82, 27.81, 27.74, 27.67, 27.64, 24.31, 23.50, 23.04, 22.93, 18.22, 12.63; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 711.6139 (100 %), cale 711.6152; ([M + Na] +) 733.5974 (46.1%), cale.733.5972.

Example 3 Compound 30: Colic acid (3.0 g, 7.3 mmol) was dissolved in CH2Cl2 (50 mL) and methanol (5 mL). Dicyclohexylcarbodiimide (DCC) (1.8 g, 8.8 mmol) was added followed by N-hydroxysuccinimide (-100 mg) and benzylmethylamine (1.1 g, 8.8 mmol). The mixture was stirred for 2 hours, and then filtered. The filtrate was concentrated and subjected to chromatography (SiO2, 3% MeOH in CH2Cl2) to give 3.0 g of a white solid (81% yield). P.f. 184-186 ° C; (SiO2, 3% MeOH in Ch2Cl2) to give 3.0 g of a white solid (81% benefit), m.p. 184-186 ° C; IR (neat) 3325, 2984, 1678 cm "1; 1 H NMR (CDCl 3, 200 MHz) d 7.21 (m, 5 H), 4.51 (m, 2 H), 3.87 (m, 1 H), 3.74 (m, 2 H) , 3.36 (m, 2H), 2.84 (s, 3H), 2.48-0.92 (series of multiplets, 28 H), 0.80 (s, 3H), 0.58 (d, J = 6.5 Hz, 3H); 3C NMR (CDCI3 , 50 MHz) d 174.30, 173.94, 137.36, 136.63, 128.81, 128.46, 127.85, 127.50, 127.18, 126.28, 72.96, 71.76, 68.35, 53.39, 50.65, 48.77, 46.91, 46.33, 41.44, 39.36, 39.18, 35.76, 35.27 , 34.76, 33.87, 31.54, 34.19, 31.07, 30.45, 28.11, 27.63, 26.14, 25.59, 24.92, 23.26, 17.51, 12.41; FAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] + ) 512 (100%) cale.512.

Compound 31: Compound 30 (2.4 g, 4.7 mmol) was added to a suspension of LiAIH (0.18 g, 4.7 mmol) in THF (50 mL). The mixture was refluxed for 24 hours, then cooled to 0 ° C. An aqueous solution of Na 2 SO 4 was carefully added until the gray color of the mixture dissipated. The salts were filtered and the filtrate was concentrated in vacuo to yield 2.1 g of a white solid (88%). The product proved to be of sufficient purity for additional reactions. P.f. 70-73 ° C; IR (neat) 3380, 2983, 1502 ern "1; 1 H NMR (CDCl 3, 300 MHz) d 7.23 (m, 5H), 3.98 (bs, 2H), 3.81 (m, 3H), 3.43 (m, 3H), 2.74 (m, 2H), 2.33 (m, 3H), 2.25 (s, 3H), 2.10-0.90 (series of multiplets, 24 H), 0.98 (s, 3H), 0.78 (s, 3H), 13C NMR ( CDCI3, 75 MHz) d 135.72, 129.63, 128.21, 128.13, 125.28, 72.91, 71.63, 62.05, 60.80, 56.79, 47.00, 46.23, 41.44, 40.81, 39.41, 35.42, 35.24, 34.63, 34.02, 33.22, 31.73, 30.17, 29.33, 29.16, 28.02, 27.49, 26.17, 25.55, 23.10, 22.48, 22.33, 17.54, 12.65; FAB-MS (thioglycerol matrix) m / e: QM + H] +) 498 (100%), cale.498.

Compound 32: Compound 31 (0.36 g, 0.72 mmol) was dissolved in CH2Cl2 (15 mL) and Bocglycine (0.51 g, 2.89 mmol), DCC (0.67 g, 3.24 mmol) and dimethylaminopyridine (DMAP) (-100 mg were added). ). The mixture was stirred under N2 for 4 hours and then filtered. After concentration and chromatography (SiO2, 5% MeOH in CH2CI2), the product was obtained as 0.47 g of a clear crystal (68%). 1 H NMR (CDCl 3, 300 Mhz) d 7.30 (m, 5 H), 5.19 (bs, 1 H), 5.09 (bs, 3 H), 5.01 (bs, 1 H), 4.75 (m, 1 H), 4.06-3.89 (m, 6H), 2.33 (m, 2H), 2.19 (s, 3H), 2.05-1.01 (series of multiplets, 26 H), 1.47 (s, 9 H), 1.45 (s, 18 H), 0.92 (s, 3H) ), 0.80 (s, J = 6.4 Hz, 3H), 0.72 (s, 3H). 13C NMR (CDCI3, 75 MHz) d 170.01, 169.86, 169.69, 155.72, 155.55, 139.90, 129.05, 128.17, 126.88, 19.86, 76.53, 75.09, 72.09, 62, 35, 57.88, 47.78, 45.23, 43.12, 42.79, 42.16 , 40.18, 37.94, 35.51, 34.69, 34.57, 34.36, 33.30, 31.31, 29.66, 28.80, 28.34, 27.22, 26.76, 25.61, 24.02, 22.83, 22.47, 17.93, 12.19; FAB-MS (thioglycerol matrix) m / e: ([M + H] +) 970 (100%), cale.970.

Compound 33: Compound 31 (0.329 g, 0.79 mmol) was dissolved in CH2Cl2 (15 mL) and Boc-β-alanine (0.60 g, 3.17 mmol), DCC (0.73 g, 3.56 mmol) and dimethylaminopyridine (DMAP) were added. (-100 mg). The mixture was stirred under N2 for 6 hours and then filtered. After concentration and chromatography (SiO2, 5% MeOH in CH2Cl2), the product was obtained as 0.58 g of a clear crystal (72%). IR (pure) 3400, 2980, 1705, 1510, cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.27 (m, 5H), 5.12 (bs, 4H), 4.93 (bs, 1H), 4.71 (m, 1H), 3.40 (m, 12H), 2.59-2.48 (m, 6H), 2.28 (m, 2H), 2.17 (s, 3H), 2.05-1.01 (series of multiplets, 26 H), 1.40 (s, 27 H), 0.90 (s, 3H), 0.77 (s, J = 6.1 Hz, 3H), 0.70 (s, 3H), 13C NMR (CDCI3, 75 MHz) d 171.85, 171.50, 171.44, 155.73, 138.62, 129.02, 128.09, 126.87, 79.18, 75.53, 74.00, 70.91, 62.20, 57.67, 47.84, 44.99, 43.28, 41.98, 40.73, 37.67, 36.12, 34.94, 34.65, 34.47, 34.20, 33.29, 31.23, 29.57, 28.74, 28.31, 28.02, 27.86, 27.12, 26.73, 25.46, 24.86, 23.95, 22.77, 22.39, 17.91, 12.14; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + H] +) 1011.6619 (100%), step 1011.6634.

Compound 6: Compound 32 (0.15 g, 0.15 mmol) was stirred with excess 4 N HCl in dioxane for 40 minutes. The dioxane and HCl were removed in vacuo leaving 0.12 g of a clear crystal (-100%). 1H NMR (CD3OD, 300 MHz) d 7.62 (bs, 2H), 7.48 (bs, 3H), 5.30 (bs, 1H), 5.11 (bs, 1H), 4.72 (bs, 1H), 4.46 (m, 1H) , 4.32 (m, 1H), 4.05-3.91 (m, 4H), 3.10 (m, 2H), 2.81 (s, 3H), 2.15-1.13 (series of multiplets, 25 H), 1.00 (s, 3H), 0.91 (bs, 3H), 0.82 (s, 3H). 13C NMR (CD3OD, 125 MHz) d 166.86, 166.50, 131.09, 130.18, 129.17, 128.55, 76.60, 75.43, 72.61, 72.04, 70.40, 66.22, 60.07, 58.00, 57.90, 54.89, 54.76, 46.44, 44.64, 43.39, 42.22 , 38.56, 36.78, 34.14, 33.92, 33.84, 31.82, 30.54, 29.67, 28.79, 27.96, 26.79, 26.00, 24.99, 23.14, 22.05, 21.82, 19.91, 17.27, 11.60; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M-4CI-3H] +) 669.4576 (100%), cale.669.4591.

Compound 7: Compound 33 (0.20 g, 0.20 mmol) was stirred with excess 4 N HCl in dioxane for 40 minutes. The dioxane and HCl were removed in vacuo leaving 0.12 g of a clear crystal (-100%). 1H NMR (CD3OD, 500 MHz) d 7.58 (bs, 2H), 7.49 (bs, 3H), 5.21 (bs, 1H), 5.02 (bs, 1H), 4.64 (m, 1H), 4.44 (m, 1H) , 4.28 (m, 1H), 3.30-2.84 (m, 14H), 2.80 (s, 3H), 2.11- 1.09 (series of multiplets, 25H), 0.99 (s, 3H), 0.89 (d, J = 4.1 Hz , 3H), 0.80 (s, 3H); 13C NMR (CD3OD, 125 MHz) d 171.92, 171.56, 171.49, 132.44, 131.32, 131.02, 130.51, 78.13, 76.61, 61.45, 57.94, 46.67, 44.80, 42.36, 40.85, 39.33, 37.03, 36.89, 36.12, 36.09, 35.79 , 35.63, 33.81, 33.10, 32.92, 32.43, 30.28, 28.43, 28.04, 26.65, 24.02, 22.86, 21.98, 18.70, 12.68; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M-4CI-3H] +) 711.5069 (43%), cale.711.5061.

Example 4 Compound 34: Diisopropyl azodicarboxylate (DIAD) (1.20 ml) 6.08 mmol) was added to triphenylphosphine (1.60 g, 6.08 mmol) in THF (100 ml) at 0 ° C and stirred for half an hour, during which time the Yellow solution became a paste. Compound 14 (2.58 g, 4.06 mmol) and p-nitrobenzoic acid (0.81 g, 4.87 mmol) were dissolved in THF (50 ml) and added to the pulp. The resulting mixture was stirred at room temperature overnight. Water (100 ml) was added and the mixture made slightly basic by adding NaHCO3 solution, followed by extraction with EtOAc (3 x 50 ml). The combined extracts were washed with brine once and dried over anhydrous Na2SO4. The desired product (2.72 g, 85% yield) was obtained as a white powder after chromatography of SiO2 (Et2O / hexanes 1: 2). P.f. 207-209 ° C; IR (KBr) 3434, 3056, 2940, 2868, 1722, 1608, 1529, 1489, 1448, 1345 cm "1; 1 H NMR (CDCl 3, 300 MHZ) d 8.30-8.25 (m, 2 H), 8.21-8.16 ( m, 2 H), 7.46-7.42 (m, 6 H), 7.31-7.18 (m, 9 H), 5.33 (bs, 1 H), 4.02 (bs, 1 H), 3.90 (bs, 1 H), 3.09-2.97 (m, 2 H), 2.68 (td, J = 14.95, 2.56 Hz, 1 H), 2.29-2.19 (m, 1 H), 2.07-1.06 (series of multiplets, 24 H), 1.01 (s) , 3 H), 0.98 (d, J = 6.6 Hz, 3 H), 0.70 (s, 3 H); 13C NMR (CDCI3, 75MHZ) d 164.21, 150.56, 144.70, 136.79, 130.77, 128.88, 127.86, 126.98, 123.70, 86.47, 73.24, 73.0, 68.70, 64.22, 47.79, 46.79, 42.15, 3976, 37.47, 35.52, 35.34 , 34.23, 33.79, 32.46, 31.12, 28.74, 27.71, 26.85, 26.30, 25.16, 23.41, 17.98, 12.77; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 808.4203 (53.8%), cale. 808.4189. The nitrobenzoate (2.75 g, 3.5 mmol) was dissolved in CH 2 Cl 2 (40 mL) and MeOH (20 mL) and 20% aqueous NaOH (5 mL) were added. The mixture was heated to 60 ° C for 24 hours. Water (100 ml) was introduced and extracted with EtOAc. The combined extracts were washed with brine and dried over anhydrous Na2SO4. The desired product 81.89 g, 85% yield) was obtained as white solid after SiO2 chromatography (3% MeOH in CH2Cl2 as a levigant). P.f. 105-106 ° C; IR (KBr) 3429, 3057, 2936, 1596, 1489, 1447, 1376, 1265, 1034, 704 cm "1; 1 H NMR (CDCl 3, 300 MHZ) d 7.46-7.42 (m, 6 H), 7.32-7.19 ( m, 9 H), 4.06 (bs, 1 H), 3.99 (bs, 1 H), 3.86 (bd, J = 2.44 hz, 1 H), 3.09-2.97 (m, 2 H), 2.47 (td, J = 14.03, 2.44 Hz, 1 H), 2.20-2.11 (m, 1 H), 2.04-1.04 (series of multiplets, 25 H), 0.97 (d, J = 6.59 Hz, 3 H), 0.94 (s, 3 H), 0.68 (s, 3 H); 13 C NMR (CDCl 3, 75 MHz) d 144.70, 128.88, 127.86, 126.97, 86.45, 73.31, 38.84, 67.10, 64.23, 47.71, 46.74, 42.10, 39.70, 36.73, 36.73, 36.15, 35.53, 35.45, 34.45, 32.46, 29.93, 28.67, 27.86, 27.71, 26.87, 26.04, 23.43, 23.16, 17394, 12.75; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] + ) 659.4064 (100%), cale.659.4076.

Compound 35: To a round bottom flask was added 34 (2.0 g, 3.14 mmol), NaH (60% in mineral oil, 3.8 g, 31.4 mmol) and THF (150 ml). The suspension was refluxed for 2 hours followed by the addition of allyl bromuor (2.72 ml, 31.4 ml). After refluxing for 28 hours, another 10 eq. of NaH and allyl bromide. After 72 hours, another 10 eq. of NaH and allyl bromide. After 115 hours, TLC showed almost no starting material or intermediates. Water (100 ml) was added to the suspension carefully, followed by extraction with EtOAc (5 x 50 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO. The desired product (1.81 g, 79% yield) was obtained as a yellowish crystal after SiO2 chromatography (5% EtOAc / hexanes). IR (pure) 3060, 3020, 2938, 2865, 1645, 11596, 1490, 1448, 1376, 1076, 705 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.46-7.42 (m, 6H), 7.31-7.18 (m, 9H), 6.06-5.85 (m, 3H), 5.35-5.20 (m, 3H), 5.15-5.06 (m, 3H), 4.10-4.00 (m, 2H), 3.93-3.90 (m, 2H) , 3.85-3.79 (ddt, J = 13.01, 4.88, 1.59 Hz, 1H), 3.73-3.66 (ddt, J = 13.01, 5.38, 1.46 Hz, 1H), 3.58 (bs, 1H), 3.54 (bs, 1H) , 3.32 (d, J = 2.93 Hz, 1H), 3..07-2.96 (m, 2H), 2.36 (td, J = 13.67, 2.68 Hz, 1H), 2.24-2.10 (m, 2H), 2.03- 1.94 (m, 1H), 1.87-0.86 (series of multiplets, 20H), 0.91 (s, 3H), 0.90 (d, J = 6.83 Hz, 3H), 0.64 (s, 3H); 3C NMR (CDCI3, 75 MHz) d 144.77, 136.29, 136.13, 128.90, 127.86, 126.94, 116.13, 115.51, 115.42, 86.44, 81.11, 75.65, 73.92, 69.40, 68.81, 64.43, 46.68, 46.54, 42.93, 39.93, 36.98, 35.66, 35.14, 32.83 , 32.54, 30.48, 28.51, 27.72, 27.64, 26.82, 24.79, 23.65, 23.43, 23.40, 18.07, 12.80; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + H] +) 757.5185 (12.9%), cale.757.5196.

Compound 36: Ozone was bubbled through a solution of 35 (0.551 g, 0.729 mmol) in CH2Cl2 (40 mL) and MeOH (20 mL) at -78 ° C until the solution turned deep blue. The excess ozone was displaced with oxygen. Methyl sulfide (1 ml) was added followed by the addition of NaBH 4 (0.22 g, 5.80 mmol) in 5% NaOH solution and methanol. The resulting mixture was stirred overnight at room temperature and washed with brine. The brine was then extracted with EtOAc (3 x 20 ml). The combined extracts were dried over Na2SO4. The desired product (0.36 g, 65% yield) was obtained as a colorless crystal after SiO2 chromatography (5% MeOH / CH2Cl2). IR (pure) 3396, 3056, 2927, 1596, 1492, 1462, 1448, 1379, 1347, 1264, 1071 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.46-7.42 (m, 6H), 7.32-7.18 (m, 9H), 3.77-3.57 (series of multiplets, 10 H), 3.48-3.44 (m, 2H), 3.36-3.30 (m, 2H), 3.26-3.20 (m, 1H), 3.04-2.99 (m , 2H), 2.37-0.95 (multi-full series, 27 H), 0.92 (s, 3H), 0.91 (d, J = 6.59 Hz, 3H), 0.67 (s, 3H); 13C NMR (CDCI3, 75 MHz) d 144.69, 128.87, 127.84, 126.94, 86.44, 81.05, 76.86, 74.65, 69.91, 69.22, 68.77, 64.24, 62.44, 62.42, 62.26, 46.92, 46.54, 42.87, 39.73, 36.86, 35.52, 35.13, 32.82, 32.54, 30.36 , 28.71, 27.61, 27.44, 26.79, 24.82, 23.51, 23.38, 23.31, 18.28, 12.74; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + Na] +) 791.4844 (96.4%), cale.791.4863.

Compound 37: NEt 3 (0.23 ml, 1.66 mmol) was added to a solution of 36 (0.364 g, 0.47 mmol) in dry CH 2 Cl 2 (30 ml) at 0 ° C under N 2 followed by the introduction of mesyl chloride (0.12 ml, 1.56 mmol). The mixture was stirred for 10 minutes and H 2 O (10 ml) was added to quench the reaction, followed by extraction with EtOAc (3 x 30 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO4. Chromatography of SiO2 (EtOAc / hexanes 1: 1) gave the desired product (0.411 g, 86% yield) as white crystal. IR (pure) 3058, 3029, 2939, 2868, 1491, 1461, 1448, 1349, 1175, 1109, 1019 cm "1; 1 H NMR (CDCl 3, 300 MHZ) d 7.46-7.42 (m, 6 H), 7.31- 7.19 (m, 9 H), 4.35-4.26 (m, 6 H), 3.84-3.74 (m, 2 H), 3.64-3.56 (m, 4 H), 3.49-3.34 (m, 3 H), 3.06 ( s, 3 H), 3.04 (s, 3 H), 3.02 (s, 3 H), 3.09-2.95 (m, 2 H), 2.28 (bt, J = 14.89 Hz, 1 H), 2.09-0.86 (series of multiplets, 21 H), 0.92 (s, 3 H), 0.90 (d, J = 6.78 Hz, 3 H), 0.66 (s, 3 H); 13C NMR (CDCI3, 75MHZ) d 144.66, 128.86, 127.86, 126.97, 86.46, 81.28, 77.18, 75.00, 70.14, 69389, 69.13, 66.49, 65.85, 65.72, 64.22, 47.06, 46.35, 42.77, 39.58, 37.81, 37.64 , 37.55, 36.75, 35.48, 35.02, 32.59, 32.52, 30.27, 28.43, 27.56, 27.52, 26.92, 24.62, 23.34, 23.25, 23.10, 18.24, 12.64; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + Na] +) 1025.4207 (100%), cale. 1025.4189.

Compound 38: The suspension of 37 (0.227 g, 0.277 mmol) and NaN3 (0.147 g, 2.27 mmol) in dry DMSO (5 ml) was stirred at 80 ° C overnight, diluted with H2O (50 ml) and extracted with EtOAc (3 x 20 ml). The extracts were washed with brine once and dried over anhydrous Na2SO. Chromatography of SiO 2 (EtOAc / hexanes 1: 8) gave the desired product (0.153 g, 80% yield) as a yellow oil. IR (pure) 2929, 2866, 2105, 1490, 1466, 1448, 1107, 705 cm "1; 1 H NMR (CDCl 3, 300 MHZ) d 7.46-7.42 (m, 6 H), 7.32-7.19 (m, 9 H ), 3.80-3.74 (m, 1 H), 3.70-3.55 (series of multiplets, 5 H), 3.41-3.19 (series of multiplets, 9 H), 3.04-2.98 (m, 2 H), 2.41 (td, J = 13.1, 2.44 hz, 1 H), 2.29-2.14 (m, 2 H), 2.04-0.86 (series of multiplets, 20 H), 0.93 (s, 3 H), 0.91 (d, J = 6.60 Hz, 3 H), 0.66 (s, 3 H), 13 C NMR (CDCl 3, 75 MHz) d 144.78, 128.93, 127.87, 126.96, 86.46, 81.30, 77.16, 75.21, 67.99, 67.44, 67.03, 64.41, 51.64, 51.57, 51.33 , 46.71, 46.30, 42.35, 39.75, 36.72, 35.64, 35.20, 32.52, 32.42, 30.17, 28.63, 27.80, 27.22, 26.90, 24.80, 23.55, 23.30, 23.24, 18.23, 12.65; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + Na] +) 866.5049 (96.9%), cale.866.5057.

Compound 39: p-Toluenesulfonic acid (1.72 mg) was added in the solution of 38 (0.153 g, 0.18 mmol) in CH2Cl2 (5 mL) and MeOH (5 mL), and the mixture was stirred for 2.5 hours. The saturated NaHCO3 solution (5 ml) was introduced followed by the introduction of brine (30 ml). The aqueous mixture was extracted with EtOAc and the combined extracts were washed with brine and dried over Na2SO. The desired product (0.10 g, 92% yield) was obtained as a pale yellowish oil after chromatography of SiO2 (EtOAc / hexanes 1: 3). IR (pure) 3426, 2926, 2104, 1467, 1441, 1347, 1107 cm "1; 1 H NMR (CDCl 3, 300 MHZ) d 3.81-3.74 (m, 1 H), 3.71-3.54 (m, 7 H), 3.41-3.19 (m, 9 H), 2.41 (td, J = 13.61, 2.32 Hz, 1 H), 2.30-2.14 (m, 2 H), 2.07-1.98 (m, 1 H), 1.94-0.95 (series of multiplets, 21 H), 0.95 (d, J = 6.35 Hz, 3 H), 0.93 (s, 3 H), 0.69 (s, 3 H); 13 C NMR (CDCI 3, 75 MHZ) d 81.22, 77.08, 75.13 , 67.94, 67.36, 66.97, 63.76, 51.59, 51.51, 51.26, 46.51, 46.24, 42.31, 39.68, 36.64, 35.58, 35.12, 32.34, 31.92, 30.11, 29.55, 28.54, 27.82, 27.16, 24.75, 23.47, 23.23, 23.18 , 18.15, 12.56; HRFAB-MS (thioglycerol + Na + matrix) m / e: QM + Na] +) 624.3966 (54.9%), cale.624.3962.

Compound 40: To a solution of 39 (0.10 g, 0.166 mmol) in CH 2 Cl 2 (8 ml) at 0 ° C was added NEt 3 (34.8 μl, 0.25 mmol) under N 2, followed by the introduction of mesyl chloride (15.5 μl, 0.199 mmol). The mixture was stirred 15 minutes, the addition of H 2 O (3 ml) and brine 820 ml) was followed by extraction with EtOAc (4 x 10 ml). The combined extracts were washed with brine once and dried over Na2SO4. After removal of the solvent, the residue was mixed with N-benzylmethylamine (0.5 ml) and heated at 80 ° C under N2 overnight. Excess N-benzylmethylamine was removed in vacuo and the residue was subjected to SiO2 chromatography (EtOAc / hexanes 1: 4) to give the product (0.109 g, 93% yield) as a yellow oil. IR (pure) 2936, 2784, 2103, 1467, 1442, 1346, 1302, 1106, 1027 ern "1; 1 H NMR (CDCl 3, 300 MHZ) d 7.32-7.23 (m, 5 H), 3.81-3.74 (m, 1 H), 3.71-3.55 (m, 5 H), 3.47 (s, 2 H), 3.41-3.74 (m, 1 H), 3.71-3.55 (m, 5 H), 3.47 (s, 2 H), 3.41-3.19 (m, 9 H), 2.46-2.11 (m, 5 H), 2.18 (s, 3 H), 2.03-0.85 (series of multiplets, 20 H), 0.93 (s, 3 H), 0.93 ( d, J = 6.35 Hz, 3 H), 0.67 (s, 3 H), 13 C NMR (CDCl 3, 75 MHz) d 139.54, 129.26, 128.32, 126.97, 81.26, 77.12, 75.17, 67.98, 67.42, 67.00, 62. 50, 58.41, 51.61, 51.54, 51.29, 46.66, 46.28, 42.46, 42.32, 39.72, 36.68, 35.76, 35.16, 33.75, 32.38, 30.15, 28.59, 27.85, 27.19, 24.77, 24.15, 23.53, 23.28, 23.22, 18.28, 12.60; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 705.4929 (100%), cale.705.4928.

Compound 8: A suspension of 40 (0.109 g, 0.155 mmol) and LiAIH4 (23.5 mg, 0.62 mmol) in THF (20 ml) was stirred under N2 overnight. Na2SO410H2O was carefully added and stirred until the gray color disappeared. Anhydrous Na2SO4 was added and the white precipitate was filtered and rinsed with dry THF. After solvent removal, the residue was dissolved in minimal CH2Cl2 and filtered. The desired product (0.091 g, 94% yield) was obtained as a colorless oil after the solvent was removed. IR (pure) 3371, 3290, 3027, 2938, 2862, 2785, 1586, 1493, 1453, 1377, 1347, 1098 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 7.31-7.21 (m, 5 H), 3.65-3.53 (m, 4 H), 3.47 (s, 2 H), 3.42-3.34 (m, 2 H), 3.30 (bs) , 1 H), 3.26-3.20 (m, 1 H), 3.14-3.09 (m, 1 H), 2.89-2.81 (series of multiplets, 29 H), 0.93 (d, J = 6.59 Hz, 3 H), 0.92 (s, 3 H), 0.67 (s, 3 H); 13C NMR (CDCI3, 75MHZ) d 139.34, 129.16, 128.24, 126.90, 80.75, 76.44, 74.29, 70.58, 69.88, 69.75, 62.47, 58.27, 46.66, 46.47, 42.75, 42.63, 42. 51, 42.35, 39.77, 36.87, 35.73, 35.04, 33.77, 32.90, 30.38, 28.71, 27.70, 27.32, 24.89, 24.09, 23.53, 23.36, 23.25, 18.24, 12.62; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 627.5199 (23.3%), cale.627.5213.

Compound 9: To a solution of 23 (0.18 g, 0.28 mmol) in dry DMF (4 ml) were added NaH (0.224 g, 60% in mineral oil, 5.60 mmol) and 1-bromo octane (0.48 ml, 2.80 mmol). The suspension was stirred under N2 at 65 ° C overnight followed by the introduction of H2O (60 ml) and extraction with ether (4 x 20 μl). The combined extracts were washed with brine and dried over Na2SO4. Chromatography of SiO2 (hexanes and 5% EtOAc in hexanes) provided the desired product (0.169 g, 80% yield) as a pale yellowish oil. I R (pure) 2927, 2865, 2099, 1478, 1462, 1451, 1350, 1264, 1 105 crrf1; 1 H NMR (CDCI3, 300 MHZ) d 3.69-3.35 (series of multiplets, 15 H), 3.26-3.02 (series of multiplets, 4 H), 2.19-2.02 (m, 3 H), 1 .97-1. 16 (series of multiplets, 37 H), 1 .12-0.99 (m, 2 H), 0.92-0.86 (m, 9 H), 0.65 (s, 3 H); 3C NMR (CDCl 3, 75 MHz) d 80.69, 79.84, 76. 1 3, 71 .57, 71. 1 5, 65.07, 64.49, 64.39, 49.08, 48.99, 48.80, 46.68, 46.45, 42.72, 42.05, 39.88, 35374, 35.49, 35.36, 35.14, 32.42, 32.03, 30.01, 29.85, 29.81, 29.76, 29.67, 29.48, 29 14, 27.92, 27.80, 27.70, 26.58, 26.42, 23.59, 23.09, 22.92, 22.86, 1 8.1 1, 14.31, 12.65; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 778.5685 (22.1%), cale. 778.5683. Triazide (0.169 g, 0.224 mmol) and L1AIH4 (0.025 g, 0.67 mmol) were suspended in anhydrous THF (10 mL) and stirred under N2 at room temperature overnight followed by careful introduction of Na2SO4 hydrate. After the gray color disappeared, anhydrous Na2SO was added and stirred. The white precipitate was removed by filtration and washed with THF. After removal of solvent, the residue was dissolved in 1 M hydrochloric acid and the aqueous solution was extracted with ether (5 ml) once. Then the aqueous solution was made basic by adding 20% aqueous NaOH solution followed by extraction with Et 2 O (4 x 5 ml). The combined extracts were washed, dried and concentrated. The residue was then subjected to SiO2 chromatography (MeOH / CH2CI2 (1: 1) followed by MeOH / CH2CI2 / NH3 H2O (4: 4: 1)) to provide the desired product (0.091 g, 60% yield) as a colorless oil. IR (pure) 3361, 2927, 2855, 1576, 1465, 1351, 1105 cm "1; 1 H NMR (CDCl 3, 300 MHz) d 4.86, (bs, 6 H), 3.77-3.72 (m, 1 H), 3.70 -3.61 (m, 1 H), 3.57-3.53 (m, 3 H), 3.43-3.38 (m, 4 H), 3.34-3.27 (m, 2 H), 3.18-3.10 (m, 2 H), 2.84 -2.71 (m, 6 H), 2.22-2.07 (m, 3 H), 2.00-1.02 (series of multiplets, 39 H), 0.97-0.88 (m, 9 H), 0.71 (s, 3 H); NMR (CDCI3, 75 MHz) d 82.20, 81.00, 77.62, 72.52, 72.06, 68.00, 67.92, 67.39, 48.20, 47.53, 44.26, 43.40, 41.42, 41.15, 40.84, 40.35, 36.88, 36.73, 36.42, 36.11, 34.24, 34.05, 33.94, 33.67, 33.17, 30.95, 30.72, 30.62, 29.81, 29.35, 28.87, 28.79, 27.51, 24.57, 23.90, 23.83, 23.44, 18.76, 14.62, 13.07; HRFAB-MS (thioglycerol matrix) m / e: ([M + H] +) 678.6133 (100%), cale.678.6149.

Compound 10: A suspension of 23 (0.126 g, 0.196 mmol) and LiAIH4 (0.037 g, 0.98 mmol) in THF (40 mL) was stirred at room temperature under N2 overnight, followed by the careful addition of Na2SO410H2O. After the gray color disappeared in the suspension, anhydrous Na 2 SO 4 was added and stirred until the organic layer became clear. The white precipitate was removed by filtration and washed with THF twice. THF was removed in vacuo, and the residue was subjected to SiO 2 chromatography (MeOH / CH 2 Cl 2 / NH 3 H 2 O (4: 4: 1)) to provide the desired product (0.066 g, 60% yield) as an oil colorless. IR (pure) 3365, 2933, 2865, 1651, 1471, 1455, 1339, 1103 cm "1; 1H NMR (CDCI3 / CD3OD 30%, 300 MHz) d 4.43 (bs, 7 H), 3.74-3.68 (m, 1 H), 3.66-3.60 (m, 1 H), 3.57-3.50 (m, 5 H), 3.34-3.25 (M, 2 H), 3.17-3.06 (M, 2 H), 2.84-2.74 (M, 6 H), 2.19-2.01 (M, 3 H), 1.97-0.96 (series of multiplets, 27 H), 0.94 (d, J = 7.2 Hz, 3 H), 0.92 (s, 3 H), 0.69 (s) , 3 H); 13C NMR (CDCI3, 75MHZ) d 80.44, 79.27, 75.77, 66.59, 66.53, 65.86, 62.51, 46.21, 45.84, 42.55, 41.53, 40.09, 39.43, 39.31, 39.02, 35.16, 34.93, 34.86, 34.57, 32.93, 32.71, 31.57, 28.66, 28.33, 27.64, 27.22, 23.04, 22.40, 22.29, 17.60 11.98; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 566.4889 (8.9% ), cale.566.4897.

Example 5 Compound 42: Compound 41 was prepared following the method reported by D.H.R. Barton, J. Wozniak, S.Z. Zard, A Short And Efficient Degradation of The Bile Acid Side Chain. Some Novel Reactions of Sulphines and Aketoesters (A Short and Efficient Degradation of the Side Chain of Bile Acid, Some Novel Reactions of Sulfins and Acetoesters), Tetrahedron, 1989, vol. 45, 3741-3754. A mixture of 41 (1.00 g, 2.10 mmol), ethylene glycol (3.52 mL, 63 mmol) and p-TsOH (20 mg, 0.105 mmol) was refluxed in benzene under N2 for 16 hours. Water was formed during the reaction by a Dean-Stark moisture trap. The cooled mixture was washed with NaHCO3 solution (50 ml) and extracted with Et2O (50 ml, 2 x 30 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO4. Removal of the solvent provided the product (1.09 g, 100%) as a white crystal. IR (pure) 2939, 2876, 1735, 1447, 1377, 1247, 1074, 1057, 1039 cm "1; 1 H NMR (CDCl 3, 300 MHZ) d 5.10 (t, J = 2.70 Hz, 1 H), 4.92 (d , J = 2.69 Hz, 1 H), 4.63-4.52 (m, 1 H), 3.98-3.80 (m, 4 H), 2.32 (t, J = 9.51 Hz, 1 H), 2.13 (s, 3 H) , 2.08 (s, 3 H), 2.05 (s, 3 H), 2.00-1.40 (series of multiplets, 15 H), 1.34-0.98 (m, 3 H), 1.20 (s, 3 H), 0.92 (s) , 3 H), 0.82 (s, 3 H), 13 C NMR (CDCl 3, 75 MHz) d 170.69, 170.63, 170.47, 111.38, 75.07, 74.23, 70.85, 64.95, 63.43, 49.85, 44.73, 43.39, 41.11, 37.37, 34.84, 34.80, 34.52, 31.42, 29.18, 27.02, 25.41, 24.16, 22.72, 22.57, 22.44, 21.73, 21.63, 13.40; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 521.3106 (38.6%), cale 521.3114 The triacetate (1.09 g, 2.10 mmol) was dissolved in MeOH (50 mL), NaOH (0.84 g, 21 mmol) was added to the solution, the suspension was then refluxed under N2. for 24 hours, then MeOH was removed in vacuo and the residue was dissolved in Et2O (100 ml) and washed with H2O, brine and then dried over Na2SO anhydrous. The desired product (0.80 g, 96% yield) was obtained as white solid after solvent removal. P.f. 199-200 ° C. IR (pure) 3396, 2932, 1462, 1446, 1371, 1265, 1078, 1055 cm "1; 1H NMR (10% CD3OD in CDCI3, 300 MHZ) d 4.08-3.83 (series of multiplets, 9 H), 3.44- 3.34 (m, 1 H), 2.41 (t, J = 9.28 Hz, 1 H), 2.22-2.10 (m, 2 H), 1.96-1.50 (series of multiplets, 12 H), 1.45-0.96 (series of multiplets , 4 H), 1.32 (s, 3 H), 0.89 (s, 3 H), 0.78 (s, 3 H), 13 C NMR (10% CD3OD in CDCl 3, 75 MHZ) d 112.11, 72.35, 71.57, 68.09, 64.54, 63.24, 49.36, 45.90, 41.48, 41.45, 39.18, 38.79, 35.29, 34.71, 34.45, 29.90, 27.26, 26.60, 23.65, 22.54, 22.44, 22.35, 13.46; HRFAB-MS (thioglycerol + Na + matrix) m / e : ([M + Na] +) 417.2622 (87.3%), cale.417.2617.

Compound 43: To a round bottom flask were added 42 (0.80 g, 2.03 mmol) and dry THF (100 ml) followed by the addition of NaH (60% in mineral oil, 0.81 g, 20.3 mmol). The suspension was refluxed under N2 for 30 minutes before the addition of allyl bromide (1.75 ml, 20.3 mmol). After 48 hours of reflux, another 10 eq. of NaH and allyl bromide. After another 48 hours, TLC did not show any intermediates. Cold water (50 ml) was added to the cooled suspension. The mixture was extracted with Et2O (60 ml, 2 x 30 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO4. Column chromatography of SiO2 (6% EtOAc in hexanes) gave the desired product (0.94 g, 90% yield) as a pale yellow oil. IR (pure) 3076, 2933, 2866, 1645, 1446, 1423, 1408, 1368, 1289, 1252, 1226, 1206, 1130, 1080, 1057 cm "1; 1 H NMR (CDCl 3, 300 MHZ) d 6.02-5.84 ( m, 3 H), 5.31-5.04 (m, 6 H), 4.12-4.05 (m, 2 H), 4.01-5.84 (m, 3 H), 5.31-5.04 (m, 6H), 4.12-4.05 (m , 2 H), 4.01-3.81 (m, 7 H), 3.70 (dd, J = 12.94, 5.62 Hz, 1 H), 3.55 (t, J = 2.56 Hz, 1 H), 3.33 (d, J = 2.93 Hz, 1 H), 3.18-3.08 (m, 1 H), 2.65 (t, J = 10.01 Hz, 1 H), 2.32-2.14 (m, 3 H), 1.84-1.45 (series of multiplets, 10 H) , 1.41-1.22 (m, 3 H), 1.27 (s, 3 H), 1.14-0.92 (m, 2 H), 0.89 (s, 3 H), 0.75 (s, 3 H); 13C NMR (CDCI3, 75 MHZ) d 136.38, 136.07, 136.00, 116.31, 115.54, 115.38, 112.34, 80.07, 79.22, 75.05, 69.83, 69.34, 68.82, 65.14, 63.24, 48.80, 45.96, 42.47, 42.15, 39.40, 35.55, 35.16, 35.15, 29.04, 28.22, 27.52, 24.21, 23.38, 23.11, 22.95, 22.58, 13.79; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 537.3549 (100%), cale.537.3556.

Compound 44: To the solution of 43 (0.94 g, 1.83 mmol) in dry THF (50 mL) was added 9-BBN (0.5 M solution in THF, 14.7 mL, 7.34 mmol) and the mixture was stirred under N2 at room temperature for 12 hours before the addition of 20% NaOH solution (4 ml) and 30% H2O2 solution (4 ml). The resulting mixture was then refluxed for one hour followed by the addition of brine (100 ml) and extracted with EtOAc (4 x 30 ml). The combined extracts were dried over anhydrous Na2SO4. After solvent removal, the residue was purified by SiO2 column chromatography (EtOAc followed by 10% MeOH in CH2Cl2) to give the product (0.559 g, 54% yield) as a colorless oil. IR (pure) 3410, 2933, 2872, 1471, 1446, 1367, 1252, 1086 cnT1; 1 H NMR (CDCl 3, 300 MHZ) d 4.02-3.52 (multiplet series, 17 H), 3.41-3.35 (m, 1 H), 3.29 (d, J = 2.44 hz, 1 H), 3.22-3.15 (m, 3 H), 2.58 (t, J = 10.01 Hz, 1 H), 2.27-1.95 (m, 3 H), 1.83-1.48 (series of multiplets, 16 H), 1.40-0.93 (series of multiplets, 5 H) , 1.27 (s, 3 H), 0.90 (s, 3 H), 0.75 (s, 3 H); 13C NMR (CDCI3, 75MHZ) d 112.41, 80.09, 79.09, 76.31, 66.70, 66.02, 65.93, 64.80, 63.26, 61.53, 61.25, 60.86, 48.59, 45.80, 42.51, 51.72, 39.10, 35.36, 35.02, 34.98, 32.87 , 32.52, 32.40, 28.88, 27.94, 27.21, 24.33, 23.02, 22.84 (2 C's), 22.44, 13.69; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 591.3881 (100%), cale.591.3873.

Compound 45: To a solution of 50 (0.559 g, 0.98 mmol) in acetone (40 ml) and water (4 ml) was added PPTS (0.124 g, 0.49 mmol) and the solution was refluxed under N2 for 16 hours. The solvent was removed under reduced pressure. Then water (40 ml) was added to the residue and the mixture was extracted with EtOAc (40 ml, 2 x 20 ml). The combined extracts were washed with brine, dried and evaporated to dryness. Column chromatography of SiO2 (8% MeOH in CH2Cl2) of the residue gave the desired product (0.509 g, 98% yield) as a clear oil. IR (pure) 3382, 2941, 2876, 1699, 1449, 1366, 1099 cm "1; H NMR (CDCl 3, 300 MHZ) d 3.83-3.72 (m, 8 H), 3.66 (t, J = 5.62 Hz, 2 H). 3.54 (bs, 2 H), 3.43-3.28 (m, 4 H), 3.24-3.12 (m, 2 H), 2.26-2.00 (m, 4 H), 2.08 (s, 3 H), 1.98 -1.50 (series of multiplets, 15 H), 1.42-0.96 (series of multiplets, 6 H), 0.90 (s, 3 H), 0.62 (s, 3 H); 13C NMR (CDCI3, 75 MHZ) d 210.49, 78.87, (2 C's), 76.30, 66.86, 66.18, 65.69, 61.74, 61.43, 60.71, 55.31, 48.05, 43.02, 41.58, 39.53, 35.28, 35.09, 34.96, 32.77, 32.70, 32.31, 31.12, 28.72, 27.88, 27.14 , 23.47, 22.75, 22.47, 22.34, 13.86; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 547.3624 (100%), cale.547.3611.

Compound 46: To a solution of 45 (0.18 g, 0.344 mmol) in dry CH 2 Cl 2 (10 mL) at 0 ° C was added Et 3 N (0.168 mL, 1.20 mmol) followed by the addition of mesyl chloride (0.088 mL, 1.13 mmol). After 10 minutes, H2O (3 ml) and brine (30 ml) were added. The mixture was extracted with EtOAc (30 ml, 2 x 10 ml) and the extracts were washed with brine and dried over anhydrous Na2SO. After solvent removal, the residue was dissolved in DMSO (5 ml) and NaN3 (0.233 g, 3.44 mmol). The suspension was heated to 50 ° C under N2 for 12 hours. H 2 O (50 ml) was added to the cold suspension and the mixture was extracted with EtOAc (30 ml, 2 x 10 ml) and the extracts were washed with brine and dried over anhydrous Na 2 SO 4. Column chromatography of SiO2 (EtOAc / hexanes 1: 5) gave the product (0.191 g, 88% yield for two steps) as a pale yellow oil. IR (pure) 2933, 2872, 2096, 1702, 1451, 1363, 1263, 1102 crrf1; 1 H NMR (CDCl 3, 300 MHZ) d 3.72-3.64 (m, 2 H), 3.55-3.24 (series of multiplets, 11 H), 3.18-3.02 (m, 2 H), 2.22-2.02 (m, 4 H) , 2.08 (s, 3 H), 1.95-1.46 (series of multiplets, 15 H), 1.38-0.96 (series of multiplets, 6 H), 0.89 (s, 3 H), 0.62 (s, 3 H); 13C NMR (CDCI3, 75MHZ) d 210.36, 79.69, 79.22, 75.98, 65.08, 64.80, 64.53, 55.31, 48.93, 48.86, 48.76, 48.06, 43.03, 41.91, 39.66, 35.44, 35.31, 35.12, 31.04, 29.77, 29.69 , 29.67, 28.99, 28.10, 27.65, 23.60, 22.99, 22.95, 22.50, 14.00; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + Na] +) 622.3820 (100%), cale. 622.3805.Compound 11: Compound 46 (0.191 g, 0.319 mmol) was dissolved in dry THF (20 ml) followed by the addition of LiAIH4 (60.4 mg, 1.59 mmol). The gray suspension was stirred under N2 at room temperature for 12 hours. Na2SO 10H2O powder was carefully added. After the gray color in the suspension disappeared, anhydrous Na2SO was added and the precipitate was filtered. After solvent removal, the residue was purified by column chromatography (silica gel, MeOH / CH2Cl2 / NH3 H2O 28% 3: 3: 1). After the majority of the solvent had been removed from the collected fractions, 5% HCl solution (2 ml) was added to dissolve the milky residue. The resulting clear solution was then extracted with Et2O (2 x 10 ml). Then the 20% NaOH solution was added until the solution became strongly basic. CH2Cl2 (20 ml, 2 x 10 ml) was used to extract the basic solution. The combined extracts were dried over anhydrous Na2SO and the solvent removal gave the desired product (0.115 g, 69% yield) as a colorless oil. From 1 H NMR it appears that this compound was a mixture of two stereoisomers in C 20 at a ratio of approximately 9: 1. The stereoisomers were not separated but used as recovered. The spectra for the most abundant isomer: IR (pure) 3353, 2926, 2858, 1574, 1470, 1366, 1102 cnT1; 1 H NMR (CDCl 3 20% in CD3OD, 300 MHz) d 4.69 (bs, 7 H), 3.76-3.69 (m, 1 H), 3.63-3.53 (m, 5 H), 3.50-3.40 (m, 1 H) , 3.29 (bs, 1 H), 3.18-3.07 (m, 2 H), 2.94-2.83 (m, 1 H), 2.81-2.66 (m, 5 H), 2.23-2.06 (m, 4 H), 1.87 -1.50 (series of multiplets, 15 H), 1.39-0.96 (series of multiplets, 6 H), 1.11 (d, J = 6.10 Hz, 3 H), 0.93 (s, 3 H), 0.75 (s, 3 H) ); 13C NMR (CDCI3 20% in CD3OD, 75 MHz) d 81.46, 80.67, 77.32, 70.65, 67.90, 67.66, 67.18, 50.32, 47.17, 43.30, 43.06, 40.74, 40.64, 40.38, 40.26, 36.31, 36.28, 35.93, 34.30 , 34.02, 33.29, 29.63, 29.31, 28.43, 26.10, 24.67, 24.09, 23.96, 23.50, 13.30 for the largest isomer; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 524.4431 (64.2%), cale.524.4427.

Example 6 Compound 47: To a solution of 23 (0.15 g, 0.233 mmol) in dry CH 2 Cl 2 (15 ml) at 0 ° C was added Et 3 N (48.8 μl, 0.35 mmol) followed by the addition of CH 3 SO 2 Cl (21.7 μl, 0.28 mmol ). The mixture was stirred for 15 minutes before H2O (3 ml) was added. The saturated NaCl solution 820 ml) was then added, and the mixture was extracted with EtOAc (40 ml, 2 x 20 ml). The combined extracts were washed with brine and dried over anhydrous Na2SO. The solvent was rotovapped off and NaBr (0.12 g, 1.17 mmol) and DMF (10 ml) were added to the residue. The suspension was heated to 80 ° C under N2 for 2 hours. DMF was removed under vacuum and the residue was chromatographed on silica (1:10 EtOAc / hexanes) to give the desired product (0.191 g, 97% yield) as a pale yellow oil. 1H NMR (CDCI3, 300 MHZ) d 3.69-3.35 (series of multiplets, 13 H), 3.28-3.02 (series of multiplets, 4 H), 2.18-2.04 (m, 3 H), 2.00-1.60 (series of multiplets , 16 H), 1.58-0.96 (series of multiplets, 11 H), 0.92 (d, J = 6.34 Hz, 3 H), 0.89 (s, 3 H), 0.66 (s, 3 H); 13C NMR (CDCI3, 75MHZ) d 80.62, 79.81, 76.08, 65.07, 64.50, 64.34, 49.03, 48.98, 48.79, 46.49, 46.46, 42.73, 42.02, 39.85, 35.47, 35.34, 35.12, 34.79, 34.72, 29.82, 29.80 , 29.74, 29.11, 27.91, 27.78, 27.69, 23.55, 23.07, 22.88, 18.10, 12.62; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M-H] +) 706.3609 (63.1%), cale.706.3591; 704.3616 (52.8%), cale.704.3611.

Compound 48: Compounds 47 (0.191 g, 0.269 mmol) and 23 (0.295 g, 0.459 mmol) were dissolved in DMF (3 ml, distilled over BaO at 6 mm Hg before use), followed by the addition of NaH (0.054). g, 60% in mineral oil). The suspension was stirred under N2 at room temperature for 24 hours. H2O (100 ml) was added to quench the excess NaH and the mixture was then extracted with Et2O (40 ml, 3 x 20 ml) and the combined extracts were washed with brine and dried over anhydrous Na2SO. The desired product (0.177 g, 52% yield based on compound 239 was obtained as a pale yellow oil after SiO2 chromatography (EtOAc / hexanes 1: 6, then 1: 2). IR (pure) 2940, 2862 , 2095, 1472, 1456, 1362, 1263, 1113 cm "1; 1H NMR (CDCI3, 300 MHZ) d 3.68-3.35 (multi-stop series, 26 H), 3.28-3.02 (multiple-cut series, 8 H), 2.20-2.04 (m, 6 H), 1.96-1.60 (series of multiplets , 30 H), 1.52-0.98 (multiplex series, 12 H), 0.91 (d, J = 6.59 Hz, 6 H), 0.89 (s, 6 H), 0.65 (s, 6 H); 13C NMR (CDCI3, 75MHZ) d 80.86, 79.83, 76.13, 71.71, 65.06, 64.48, 64.39, 49.08, 48.98, 48.80, 46.64, 46.44, 42.71, 42.04, 39.88, . 73, 35.49, 35.36, 35.14, 32.41, 29.84, 29.81, 29.76, 29.14, 27.92, 27.78, 27.69, 26.58, 23.59, 23.08, 22.92, 18.12, 12.64.

Compound 12: Compound 48 (0.219 g, 0.173 mmol) was dissolved in dry THF (10 mL) followed by the addition of LiAIH (65 mg, 1.73 mmol). The gray suspension was stirred under N2 at room temperature for 12 hours. The Na2SO 10H2O powder was carefully added. After the gray color disappeared in the suspension, anhydrous Na2SO4 was added and the precipitate was filtered. After solvent removal, the residue was purified by column chromatography (silica gel, MeOH / CH 2 Cl 2/28% NH 3 H 2 O 2.5: 2.5: 1). After the majority of the solvent was removed from the collected fractions, a 5% HCl solution (2 ml) was added to dissolve the milky residue. The resulting clear solution was then extracted with Et2O (2 x 10 ml). Then a 20% NaOH solution was added until the solution became strongly basic. CH2Cl2 (20 ml, 2 x 10 ml) was used to extract the basic solution. The combined extracts were dried over anhydrous Na2SO and the solvent removal gave the desired product (0.147 g, 76% yield) as a white crystal. IR (pure) 3364, 3287, 2934, 2861, 1596, 1464, 1363, 1105 cm "1; 1H NMR (CDCl320% in CD3OD, 300 MHz) d 4.74 (bs, 12 H), 3.75-3.70 (m, 2 H), 3.65-3.61 (m, 2 H), 3.57-3.52 (m, 6 H), 3.40 (t, J = 3.60 Hz, 4 H), 3.30 (bs, 4 H), 3.16-3.10 (m, 4 H), 2.84-2.73 (m, 12 H), 2.18-2.07 (m, 6 H), 1.97-1.61 (series of multiplets, 30 H), 1.58-0.98 (series of multiplets, 24 H), 0.95 ( d, J = 6.84 Hz, 6 H), 0.94 (s, 6 H), 0.70 (s, 6 H), 13 C NMR (CDCI320% in CD3OD, 125 MHZ) d 81.70, 77.09, 72.34, 67.75 (2 C's) , 67.07, 47.80, 47.13, 43.76, 42.87, 41.20, 40.65, 40.58, 40.14, 36.43, 36.25, 36.08, 35.77, 34.15, 33.87 (2 C's), 33.18, 29.55, 28.92, 28.47, 28.42, 27.25, 24.27, 23.54 , 23.41, 18.70, 13.07; HRFAB-MS (thioglycerol + Na + matrix) m / e: ([M + H] +) 1 1 1 3.9625 (68.8%), cale 1 1 1 3.961 0.

Example 7 Test of compounds with gram-negative bacteria Measurements of MIC and M BC General: Tryptic soy broth (TSB) was made by dissolving 27.5 grams of tryptic soy broth without dextrose (DIFCO Laboratories) in 1 liter of deionized water and sterilizing at 1 21 ° C for 1 5 minutes. Solid agar plates (TSA) were made by dissolving 6.4 grams of tryptic soy broth and 1.2 grams of agar (purified grade, Fischer Scientific) in 800 ml of deionized water and sterilizing at 1 21 ° C for 20 minutes. Aliquots (20 ml) of the homogeneous solution were then drained in sterile plastic petri dishes (1 00 x 15 mm, Fisher Scientific). The solutions of compounds 1-12 were made by dissolving the HCl salt of the respective compound in an appropriate amount of deionized and sterilized water followed by microfiltration.

Representative procedure for measuring MIC and M BC values: A suspension of E. coli (ATCC 1 0798) containing -1 06 CFU (colony forming units) / ml was prepared from a culture incubated in TSB at 37 ° C for 24 hours. Aliquots of 1 ml of the suspension were added to test tubes containing 1 ml of TSB and concentrations that vary in increments of 1 -1 2 and / or erythromycin or novobiocin. In the sensitization experiments, erythromycin or novobiocin were added 1 5 minutes after 1-12. The samples were subjected to stationary incubation at 37 ° C for 24 hours. The sample turbidity was determined by measuring the absorption at 760 nm (HP 8453 UV-Visible Chemstation, Hewlett Packard). Additionally, an aliquot of each sample that does not show measurable turbidity was subcultured on TSA plates (the aliquots were diluted to provide less than 300 CFU). The colonies that grew in the subculture after the overnight incubation were counted and the number of CFU / ml in the samples was calculated. The calculated values were compared with the number of CFU / ml in the original inoculum. MIC values were determined as the concentrations of the compounds studied in which the number of CFU / ml remained constant or decreased after incubation for 24 hours. The MBC values were determined as the lowest concentrations of the compounds studied that allowed less than 0.1% of the original bacterial suspension to survive.

Results The stereochemistry of the steroid skeleton results in different activity of the cholic acid derivatives (compare 2 and 8, Tables 1, 2, 6 and 7). The guanidino groups attached to the steroid provide lower MIC values than the compounds containing amino groups (compare 1, 2, 4 and 5, compare Tables 1 - 8). The length of the binding between the amino or guanidino groups and the steroid skeleton also influences the activity (compare 1 -3, Tables 1, 2, 6 and 7). The ester bonds between the amino groups and the steroid skeleton provide compounds with MIC values that are greater than the corresponding compounds containing ether bonds (compare 1, 2, 6 and 7, Tables 1 and 2). The group attached to the skeleton at C-20 or C-24 also influences the activity of the cholic acid derivatives. A long carbon chain attached to the steroid via an ether link at C-24 decreases the MI C of the compound as compared to the compound with a hydroxyl group at C-24 (compare 2, 9 and 1 0, Tables 1, 2, 6 and 7). The short chains of carbon or oxygen linked in C-20 decrease the MIC values of the cholic acid derivatives (compare 10 and 11, Tables 1 and 2). Covalently linking the cholic acid derivatives increases the activity of the compounds (compare 1 0 and 12, Tables 1 and 2).

Table 1 Measurement of MIC and MBC values of 1-12 with E. coli (ATCC 10798) Value not measured.

Table 2 Measurement of the 1-12 concentrations required to decrease the MIC of erythromycin from 70 μg / ml to 1 μg / ml with E. coli (ATCC 10798) Value not measured.

Table 3 Measurement of the concentrations of 1, 2, 4 and 5 required to reduce the Novobiocin MIC from > 500 μg / ml at 1 μg / ml with £. coli (ATCC 10798).

Value not measured.

Table 4 Measurement of MIC and MBC values of 1, 2, 4 and 5 with E. coli (ATCC 25922).

Table 5 Measurement of the concentrations of 1, 2, 4 and 5 required to decrease the M Erythromycin IC from 60 μg / ml to 1 μg / ml with E. coli (ATCC 25922).

Table 6 Measurement of MIC and MBC values of 1 -5, 8-12 with P. aureginosa (ATCC 27853).

Value not measured.

Table 7 Measurement of the concentrations of 1 -5, 8-1 2 required to decrease the MIC of erythromycin from 240 μg / ml to 5 μg / ml with P. aureginosa (ATCC 27853).

Value not measured.

Table 8 Measurement of the concentrations of 1, 2, 4 and 5 required to decrease the MIC of novobiocin from > 500 μg / ml at 1 μg / ml with P. aureginosa (ATCC 27853) EXAMPLE 8 Demonstration of Membrane Breaking Properties of Colic Acid Derivatives Using a technique described by J. M. Shupp, S. E. Travis, L. B. Price, R. F. Shand, P. Keim, Rabid Bacterial Permeabilization Reagent Useful for Enzyme Assays (Rabbit bacterial permeabilization reagent useful for enzyme assays), Biotechniques, 1 995, vol. 1 9, 1 8-20, we have shown that cholic acid derivatives increase the permeability of the outer membrane of Gram-negative bacteria. The values for mean maximum luminescence (indicating the permeabilization of the outer membrane that allows the luciferin to enter the cell) for 2 is 7 μg / ml and for 10 is 33 μg / ml. These values correspond to the M ICs measured of 2 and 1 0.

Example 9 The R groups correspond to the R groups correspond to side chain of any combila side chain of any nation of amino acids (D or L) combination of amino acids Ateaxnesenla estereoquínica dertro (oamoin example) deesteroid (uránón deanloABenesfe caso) Estemas desate anfes can be used for this trarefotrnación. (here is an example) Aryan AB in this case) Schemes of these arts can be used for this transíamadón.

Atera ± resenel number of groups (for example) hydraxtoend esterotie (0B-12in this case) Schemes untie art can be used for this transfamaiíri.

Ateraxnesenotrans for this transfofma i n Descriptions of the steroid onset materials shown above can be found in Dictionary of Steroids, Hill, R. A.; Kirk, D. N.; Makin, H. L.J.; Murphy, G. M. eds. , Chapman and Hall: New York, 1 991.

Other modalities All the features described in this specification can be mixed in any combination. Each characteristic described in this specification can be replaced by an alternative feature that serves the same purpose, equivalent or similar. Thus, unless expressly stated otherwise, each feature described is only an example of a generic series of equivalent or similar features. From the above description, one skilled in the art can find out the essential characteristics of the present invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various uses and conditions. For example, the salts, esters, ethers and amides of the novel steroid compounds described herein are within the scope of this invention. In this way, other embodiments are also within the claims.

Claims

CLAIMS 1. A compound according to formula I wherein: the fused rings A, B, C and D are independently saturated or fully or partially unsaturated; and R 1 to R 4, R 6, R, R 11, R 2, R 15, R 1 and R 17 is each independently selected from the group consisting of hydrogen, hydroxyl, a (C 1 -C 10) substituted or unsubstituted alkyl, (C 1 -C 10) ) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl , oxo, a linker group attached to a second steroid, a substituted or unsubstituted (C1-C10) aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N- HC (Q5) -C (O) -O, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10), azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O-, (C1-C10) guanidinoalkyl oxy, and (C1-C10) guanidinoalkyl carboxy, where Q5 is a side chain of any amino acid, PG It is a protective group. amino, and Rs, Re, R9, R10, R13 and R14 is each independently: deleted when one of the fused rings A, B, C or D is unsaturated in order to complete the valence of the carbon atom at that site, or selected from the group consisting of hydrogen, hydroxy, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo, a linker group attached to a second steroid, a (C1-C10) substituted or unsubstituted aminoalkyloxy , a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O-, H2N-HC (Q5) -C (O ) -N (H) -, (C1-C10) azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O, (C1-C10) guanidinoalkyloxy and (C1-C10) guanidinoalquilcarboxi, where Q5 is a side chain of any kind er amino acid, P.G. is an amino protecting group, and provided that at least two of R ^ to R14 are independently selected from the group consisting of a substituted or unsubstituted (C1-C10) aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a (C1-C10) substituted or unsubstituted aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O-, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10) azidoalkyloxy , (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O-, (C1-C10) guanidinoalkyloxy and (C1-C10) guanidinoalkylcarboxy; or a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein at least one of the following pairs is deleted and the valence of the ring carbon atoms in these deleted positions is completed with a double bond: R5 and R9; Rs and? >; and R13 and? 4. 3. The compound of claim 1, wherein at least three of Ri to R14 are independently selected from the group consisting of a substituted or unsubstituted (C1-C10) aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a (C1-C10) substituted or unsubstituted amioalkylaminocarbonyl, H2N-HC (15) -C (O) -O-, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10) azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O-, (C1-C10) guanidinoalkyloxy and (C1-C10) guanidinoalkylcarboxy. 4. The compound of claim 3, wherein the R 3 (a R 14 independently selected from the group consisting of a substituted or unsubstituted (C 1 -C 10) aminoalkyloxy, a substituted or unsubstituted (C 1 -C 10) aminoalkylcarboxy, a (C1-C10) substituted or unsubstituted aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O-, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10) azidoalkyloxy , (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O-, (C1-C10) guanidinoalkyloxy and (C1-C10) guanidinoalkylcarboxy, have the same configuration. claim 1, wherein the second steroid is a compound of formula I. The compound of claim 1, wherein the linker group is (C1-C10) alkyl-oxy- (C1-C10) alkyl. 7. The compound of claim 1, wherein: none of R5, R8, Rg, R3 and R4 is deleted. The compound of claim 1, wherein: each of R3, R7 and R? 2 is independently selected from the group consisting of a substituted or unsubstituted (C1-C10) aminoalkyloxy, a substituted (C1-C10) aminoalkylcarboxy or unsubstituted, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O, H2N-HC (Q5) -C (O) -N (H) -, (C1 Guanidinoalkyloxy, and (C 1 -C 10) cyanoalkylcarboxy, PG-HN-C (Q 5) -C (O) -O-, (C 1 -C 10) guanidinoalkyloxy and (C 1 -C 10) guanidinoalkylcarboxy, where Q 5 is a side chain of any amino acid, PG it is a protective amino group; or a pharmaceutically acceptable salt thereof. 9. The compound of claim 7, wherein: i, R ?, R4, Rs, Re, R1, R10, R11, R13, R4, R15 and R are hydrogen. The compound of claim 8, wherein: R17 is -CR18R19R2o, wherein each of R? 8, R19 and R2o, is independently selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted alkyl or unsubstituted, (C1-C10) substituted or unsubstituted hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, (C1- C10) haloalkyl, (C2-C6) alkenyl, (C2-C6) alkynyl, oxo and a linker group attached to a second steroid. The compound of claim 7, wherein: each of R3, R7 and R12 is independently selected from the group consisting of -O- (CH2) n-NH2, -O-CO- (CH2) -n- NH2, -O- (CH2) n- NH-C (NH) -NH2, -O- (CH2) n-N3, -O- (CH2) n-CN, where n is 1 to 3 and -OC (O ) -HC (Q5) -N H2, where Q5 is a side chain of any amino acid. The compound of claim 7, wherein: each of R3, R7 and R12, is -O- (CH2) n-NH2, wherein n is 1 to 4. The compound of claim 7, in where: each of R3, R7 and R1 2, is independently selected from the group consisting of -O-OC- (CH2) n-NH2, where n is 1 or 2. 14. The compound of claim 7, wherein : each of R3, R7 and R12 is independently selected from the group consisting of -O- (CH2) n-NH-C (NH) -NH2, where n is 1 to 3. The compound of claim 7 , wherein: each of R3, R7 and R? 2, is independently selected from the group consisting of -OC (O) -HC (Q5) -NH2, where Q5 is a side chain of any amino acid. 1 6. The compound of claim 1, having the following formula: The compound of claim 1, having the following formula: n = 2 Figure 2. 18. The compound of claim 1, having the formula: 19. The compound of claim 1, which has the formula: 20. The compound of claim 1, which has the formula: 10 Figure 5. 21. The compound of claim 1, having the formula: 22. The compound of claim 1, which has the formula: 23. The compound of claim 1, which has the formula: Figure 8. 24. A method for preparing the compound according to the formula wherein: the fused rings A, B, C and D are independently saturated or fully or partially unsaturated; and Ri to R4, R6, R, R11, R12, R15, R and 17 is each independently selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl , (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo , a linker group attached to a second steroid, a substituted or unsubstituted (C1-C10) aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC ( Q5) -C (O) -O, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10), azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5 ) -C (O) -O-, (C1-C10) guanidinoalkyl oxy, and (C1-C10) guanidinoalkyl carboxy, where Q5 is a side chain of any amino acid, P.G. is an amino protecting group, and R5, e, R9, R10, 13 and 14 is each independently: deleted when one of the fused rings A, B, C or D is unsaturated in order to complete the valency of the carbon at that site, or selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo, a linker group attached to a second steroid, a (C1-C10) substituted or unsubstituted aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O-, H2N-HC (Q5 ) -C (O) -N (H) -, (C1-C10) azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O, (C1-C10) guanidinoalkyloxy and (C1- C10) guanidinoalquilcarboxi, where Q5 is a c lateral adena of any amino acid, P.G. is an amino protecting group, and at least two of R to R? are independently selected from the group consisting of a substituted or unsubstituted aminoalkyloxy (C1-C10); the method comprising: contacting a compound of formula IV, wherein at least two of Ri to R 4 are hydroxyl, and the remaining portions in the fused rings A, B, C and D are defined by formula I, with an electrophile to produce an alkyl ether compound of formula IV, in where at least two of Ri to R? are (C1-C10) alkyloxy; converting the alkyl ether compounds to a precursor amino compound, wherein at least two of Ri to R14 are independently selected from the group consisting of (C1-C10) azidoalkyloxy and (C1-C10) cyanoalkyloxy; and reducing the precursor amino compound to form a compound of formula I. 25. The method of claim 19, wherein the electrophile is allyl bromide. 26. A method for producing a compound of formula I: wherein: the fused rings A, B, C and D are independently saturated or fully or partially unsaturated; and Ri to R, R6, R, R11, R? 2, R15, R and R17 is each independently selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) ) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl , oxo, a linker group attached to a second steroid, a substituted or unsubstituted (C1-C10) aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N- HC (Q5) -C (O) -O, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10), azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O-, (C1-C10) guanidinoalkyl oxy, and (C1-C10) guanidinoalkyl carboxy, where Q5 is a side chain of any amino acid, PG is an amino protecting group, and R5, R1, R9, R10, R13 and R4 are each independently: deleted when one of the fused rings A, B, C or D is unsaturated in order to complete the valency of the atom carbon at that site, or selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) substituted or unsubstituted aminoalkyl, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo, a linker group attached to a second steroid, a (C1-C10) ) substituted or unsubstituted aminoalkyloxy, a substituted or unsubstituted (C1-C10) aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O-, H2N-HC ( Q5) -C (O) -N (H) -, (C1-C10) azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O, (C1-C10) guanidinoalkyloxy and (C1-C10) guanidinoalquilcarboxi, where Q5 is a cad side of any amino acid, P.G. is an amino protecting group, and at least two of R ^ to R1 are (C1-C10) guanidoalkyloxy; the method comprising: contacting a compound of formula IV, IV wherein at least two of RT to R14 are hydroxyl, and the remaining portions in the fused rings A, B, C and D are defined by formula I, with an electrophile to produce an alkyl ether compound of formula IV, in where at least two of Ri to R? are (C1-C10) alkyloxy; converting the alkyl ether compound to a precursor amino compound, wherein at least two of Ri to R are independently selected from the group consisting of (C1-C10) azidoalkyloxy and (C1-C10) cyanoalkyloxy; reducing the precursor amino compound to produce an aminoalkyl ether compound, wherein at least two of R i to R 1 are (C 1 -C 10) aminoalkyloxy; and contacting the aminoalkyl ether compound with a guanidino producing electrophile to form a compound of formula I. 27. The method of claim 25, wherein the guanidino producing electrophile is HSO3-C (NH) -NH2. 28. A method for preparing the compound according to formula I wherein: the fused rings A, B, C and D are independently saturated or fully or partially unsaturated; and Ri to R4, R6, R7, R11, R? 2, R15, R and R17 is each independently selected from the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a substituted or unsubstituted (C1-C10) aminoalkyl, a substituted or unsubstituted aryl , C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo, a linker group attached to a second steroid, a substituted or unsubstituted (C1-C10) aminoalkyloxy, a (C1-C10) substituted or unsubstituted aminoalkylcarboxy , a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O, H2N-HC (Q5) -C (O) -N (H>, (C1-C10) , azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O-, (C1-C10) guanidinoalkyloxy, and (C1-C10) guanidinoalkyl carboxy, where Q5 is a side chain of any amino acid, PG is an amino protecting group, and R5, Rβ, Rg, R10, R13 and R? is each independently: deleted when one of the fused rings A, B, C or D is unsaturated in order to complete the valence of the carbon atom in that site, or select of the group consisting of hydrogen, hydroxyl, a (C1-C10) substituted or unsubstituted alkyl, (C1-C10) hydroxyalkyl, (C1-C10) alkyloxy- (C1-C10) alkyl, a (C1-C10) aminoalkyl substituted or unsubstituted, a substituted or unsubstituted aryl, C1-C10 haloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, oxo, a linker group attached to a second spheroid, a (C1-C10) substituted or unsubstituted aminoalkyloxy, a (C1-C10) substituted or unsubstituted aminoalkylcarboxy, a substituted or unsubstituted (C1-C10) aminoalkylaminocarbonyl, H2N-HC (Q5) -C (O) -O-, H2N-HC (Q5) -C (O) -N (H) -, (C1-C10) azidoalkyloxy, (C1-C10) cyanoalkyloxy, PG-HN-C (Q5) -C (O) -O, (C1-C10) guanidinoalkyloxy and (C1-C10) guanidinoalkylcarboxy , where Q5 is a side chain of any amino acid, PG is an amino protecting group, and at least two of R ^ to R14 are H2N-HC (Q5) -C (O) -O-, where Q5 is the side chain of any amino acid; the method comprising: contacting a compound of formula IV, where at least two of R ^ to R14 are hydroxyl, and the remaining portions in the fused rings A, B, C and D are defined by formula I, with a protected amino acid to produce a protected amino acid compound of formula IV, where at least two of Ri to R14 are (PG-HN-HC (Q5) -C (O) -O-), where Q5 is the side chain of any amino acid and PG it is a protective amino group; and removing the protecting group of the protected amino acid compound to form a compound of formula I. 29. A pharmaceutical composition comprising an effective amount of a compound of claim 1. 30. The pharmaceutical composition of claim 23, wherein the composition includes additional antibiotics. 31 A method for treating a microbial infection of a host by administering to the host an effective amount of an antimicrobial composition comprising a compound according to claim 1. 32. The method of claim 31, wherein the host is a human. 33. The method of claim 31, wherein the antimicrobial composition further comprises a second anti-microbial substance to be delivered to a microbial cell. 34. The method of claim 33, wherein the second anti-microbial substance is an antibiotic. 35. The method of claim 31, or claim 32 or claim 33, wherein the infection is a bacterial infection. 36. The method of claim 35, wherein the infection is an infection of gram-negative bacteria. 37. The method of claim 36, wherein the bacterial infection is an infection with a bacterium characterized by an outer membrane comprising a substantial percentage of lipid A. 38. A method for enhancing cellular permeability by admiring to the cell an enhancing amount of permeability of the compound of claim 1. 39. The method of claim 38 further comprising administering to the cell a substance to be introduced into the cell. 40. The method of claim 39, in which the cell is a bacterium. 41 The method of claim 40, wherein the bacterium is a gram-negative bacterium. 42. The method of claim 40, wherein the bacterium is characterized by an outer membrane comprising a substantial percentage of lipid A. 43. The method of claim 38, wherein the cell is a sperm cell and the compound is part of a spermicidal composition. 44. A method for identifying effective compounds against a microbe comprising administering a candidate compound and a compound according to claim 1 to the microbe and determining whether the candidate compound has a static or toxic effect on the microbe. 45. The method of claim 44, wherein the microbe is a bacterium. 46. The method of claim 45, wherein the bacterium is a gram-negative bacterium. 47. The method of claim 45, wherein the bacterium is characterized by an outer membrane comprising a substantial percentage of lipid A. 48. A method of controlling microbial growth comprising contacting a microbe with an effective amount of an antimicrobial composition comprising a compound according to claim 1. 49. The method of claim 48, wherein the antimicrobial composition further comprises an anti-microbial substance. 50. The method of claim 49, wherein the substance is a disinfectant, an antibiotic, a disinfectant. 51 A composition of matter comprising the compound of claim 1, in combination with a substance to be introduced into a cell. 52. The composition of claim 51, wherein the substance to be introduced into a cell is an anti-microbial substance. 53. The composition of claim 51, wherein the compound and the substance are mixed with a pharmaceutically acceptable carrier. 54. The composition of claim 52, wherein the antimicrobial substance is an antibiotic.