WO2011085454A1 - Gp120 -binding benzene compounds and saccharide compounds - Google Patents
Gp120 -binding benzene compounds and saccharide compounds Download PDFInfo
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- WO2011085454A1 WO2011085454A1 PCT/BE2011/000001 BE2011000001W WO2011085454A1 WO 2011085454 A1 WO2011085454 A1 WO 2011085454A1 BE 2011000001 W BE2011000001 W BE 2011000001W WO 2011085454 A1 WO2011085454 A1 WO 2011085454A1
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- benzoate
- pentakis
- phenylmethoxy
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- 0 *[C@@](C(C1O)O)OC(CO)[C@@]1O[C@](C(C1O)O)OC(CO)[C@]1O Chemical compound *[C@@](C(C1O)O)OC(CO)[C@@]1O[C@](C(C1O)O)OC(CO)[C@]1O 0.000 description 14
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H13/00—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids
- C07H13/02—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids
- C07H13/08—Compounds containing saccharide radicals esterified by carbonic acid or derivatives thereof, or by organic acids, e.g. phosphonic acids by carboxylic acids having the esterifying carboxyl radicals directly attached to carbocyclic rings
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C215/00—Compounds containing amino and hydroxy groups bound to the same carbon skeleton
- C07C215/46—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups bound to carbon atoms of at least one six-membered aromatic ring and amino groups bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton
- C07C215/48—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups bound to carbon atoms of at least one six-membered aromatic ring and amino groups bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton with amino groups linked to the six-membered aromatic ring, or to the condensed ring system containing that ring, by carbon chains not further substituted by hydroxy groups
- C07C215/50—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups bound to carbon atoms of at least one six-membered aromatic ring and amino groups bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton with amino groups linked to the six-membered aromatic ring, or to the condensed ring system containing that ring, by carbon chains not further substituted by hydroxy groups with amino groups and the six-membered aromatic ring, or the condensed ring system containing that ring, bound to the same carbon atom of the carbon chain
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C215/00—Compounds containing amino and hydroxy groups bound to the same carbon skeleton
- C07C215/74—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton
- C07C215/76—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton of the same non-condensed six-membered aromatic ring
- C07C215/78—Compounds containing amino and hydroxy groups bound to the same carbon skeleton having hydroxy groups and amino groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton of the same non-condensed six-membered aromatic ring containing at least two hydroxy groups bound to the carbon skeleton
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C217/00—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton
- C07C217/54—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having etherified hydroxy groups bound to carbon atoms of at least one six-membered aromatic ring and amino groups bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton
- C07C217/56—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having etherified hydroxy groups bound to carbon atoms of at least one six-membered aromatic ring and amino groups bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton with amino groups linked to the six-membered aromatic ring, or to the condensed ring system containing that ring, by carbon chains not further substituted by singly-bound oxygen atoms
- C07C217/58—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having etherified hydroxy groups bound to carbon atoms of at least one six-membered aromatic ring and amino groups bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings of the same carbon skeleton with amino groups linked to the six-membered aromatic ring, or to the condensed ring system containing that ring, by carbon chains not further substituted by singly-bound oxygen atoms with amino groups and the six-membered aromatic ring, or the condensed ring system containing that ring, bound to the same carbon atom of the carbon chain
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C217/00—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton
- C07C217/78—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having amino groups and etherified hydroxy groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton
- C07C217/80—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having amino groups and etherified hydroxy groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton having amino groups and etherified hydroxy groups bound to carbon atoms of non-condensed six-membered aromatic rings
- C07C217/82—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having amino groups and etherified hydroxy groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton having amino groups and etherified hydroxy groups bound to carbon atoms of non-condensed six-membered aromatic rings of the same non-condensed six-membered aromatic ring
- C07C217/84—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having amino groups and etherified hydroxy groups bound to carbon atoms of six-membered aromatic rings of the same carbon skeleton having amino groups and etherified hydroxy groups bound to carbon atoms of non-condensed six-membered aromatic rings of the same non-condensed six-membered aromatic ring the oxygen atom of at least one of the etherified hydroxy groups being further bound to an acyclic carbon atom
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
- C07C235/42—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton
- C07C235/44—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring
- C07C235/50—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by nitrogen atoms not being part of nitro or nitroso groups
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C235/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms
- C07C235/42—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton
- C07C235/44—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring
- C07C235/58—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring with carbon atoms of carboxamide groups and singly-bound oxygen atoms, bound in ortho-position to carbon atoms of the same non-condensed six-membered aromatic ring
- C07C235/60—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by oxygen atoms having carbon atoms of carboxamide groups bound to carbon atoms of six-membered aromatic rings and singly-bound oxygen atoms bound to the same carbon skeleton with carbon atoms of carboxamide groups and singly-bound oxygen atoms bound to carbon atoms of the same non-condensed six-membered aromatic ring with carbon atoms of carboxamide groups and singly-bound oxygen atoms, bound in ortho-position to carbon atoms of the same non-condensed six-membered aromatic ring having the nitrogen atoms of the carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
Definitions
- the present invention provides for novel compounds and for the use of said compounds for binding, titration (quantification), removing, purifying or separating the glycoprotein gp120, gp120 comprising viruses or cells infected with gp120 comprising viruses.
- the invention also provides for a method for the detection, binding, titration (quantification), removal, purification or separation of gp120, gp120 comprising viruses or cells infected with gp120 comprising viruses.
- the invention thus also relates to a separation matrix for affinity chromatography of gp120, which matrix comprises the compounds of the invention.
- the invention further provides for the use of the compounds and for methods using the compounds for directing anti-viral drugs or other agents to gp120 comprising viruses and gp120 comprising virus-infected cells.
- the present invention also provides processes for the preparation of said novel compounds.
- Gp120 is a glycoprotein exposed on the surface of the HIV envelope.
- the 120 in its name comes from its molecular weight of 120 kilodaltons.
- gp120 is essential for virus entry into cells as it plays a vital role in seeking out specific cell surface receptors for entry.
- the gp120 gene, env is around 1.5 kb long and codes for around 500 amino acids.
- Three gp120s, bound as heterotrimers to a transmembrane glycoprotein, gp41, are thought to combine in a trimer to form the envelope spike, which is involved in virus-cell attachment.
- the Human Immunodeficiency Virus can mutate frequently to stay ahead of the immune system. There is however a highly conserved region in the virus genome near its receptor binding site.
- the glycoprotein gp120 is anchored to the viral membrane, or envelope, via non-covalent bonds with the transmembrane glycoprotein, gp41. It is involved in entry into cells by binding to CD4 receptors, particularly helper T-cells and macrophages. Binding to CD4 is mainly electrostatic although there are van der Waals interactions and hydrogen bonds.
- gp120 plays a vital role in the ability of HIV-1 to enter CD4 + cells, its evolution is of particular interest. Many neutralizing antibodies bind to sites located in variable regions of gp120. The diversity of env has been shown to increase by 1-2% per year in HIV-1 group M and the variable units are notable for rapid changes in amino acid sequence length. Increases in gp120 variability result in significantly elevated levels of viral replication, indicating an increase in viral fitness in individuals infected by diverse HIV-1 variants. Further studies have shown that variability in potential N-linked glycosylation sites (PNGSs) also result in increased viral fitness.
- PNGSs N-linked glycosylation sites
- PNGSs allow for the binding of long-chain carbohydrates to the high variability regions of gp120, so it is possible that the number of PNGSs in env might affect the fitness of the virus by providing more or less sensitivity to neutralizing antibodies.
- the presence of large carbohydrate chains extending from gp120 might obscure possible antibody binding sites.
- the relationship between gp120 and neutralizing antibodies is an example of Red Queen evolutionary dynamics.
- Continuing evolutionary adaptation is required for the viral envelope protein to maintain fitness relative to the continuing evolutionary adaptations of the host immune neutralizing antibodies, and vice-versa, forming a coevolving system.
- gp120 Since CD4 receptor binding is the most obvious step in HIV infection, gp120 was among the first targets of HIV vaccine research. Efforts to develop HIV vaccines targeting gp120, however, have been hampered by the chemical and structural properties of gp120, which make it difficult for antibodies to bind to it. gp120 can also easily be shed from the surface of the virus and captured by T cells due to its loose binding with gp41. Also research is being performed to identify a new vaccine based on antibodies that hydrolyze or cleave a part of the gp120 protein, rendering it incapable of binding to lymphocytes. This binding is the first step in the process of HIV infection. The protein gp120 is necessary during the initial binding of HIV to its target cell.
- gp120 is a large glycoprotein for which the use of a gp120 binder could be helpful in the purification of gp120. Gp120 could also be targeted in order to identify, bind, purify or separate gp120 comprising viruses or cells infected with gp120 comprising viruses, such as HIV.
- compounds can be used for directing anti-viral drugs or other agents to gp120 comprising viruses.
- the present invention relates to novel gp120 binding compounds.
- the compounds have been found to bind to gp120 and can therefore be used for binding, titration (quantification, removal, purification or separation of gp120.
- the compounds can also be used to titrate (quantify), remove, detect, bind to, purify or separate gp120 comprising viruses or cells infected with gp120 comprising viruses, such as HIV.
- the compounds can furthermore be used for directing or targeting anti-viral drugs or other agents to gp120 comprising viruses or to gp120 comprising virus-infected cells.
- the first aspect of the present invention provides novel compounds which have a central benzene scaffold or a central carbohydrate scaffold, with a general structure according to the formulas (A), (B), (C), (D) or (E),
- n is independently selected from 1 and 2;
- - L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene,
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
- heteroalkynylene and (1 ,4-piperazinylene);
- each of R 1a , R 1b and R 1c is independently selected from methyl; and ethyl;
- each of R, R 3 , R 4 and R 5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; dihydroxybenzyl; trihydrozybenzyl; dimethoxyphenyl; trimethoxyphenyl; dimethoxybenzyl;
- trimethoxybenzyl and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3 and wherein each Y is independently selected from hydrogen, methyl or benzyl; or
- X-R 3 , X-R 4 or X-R 5 is a linking moiety selected from Ci-iooheteroalkyl; Ci-iooheteroalkenyl; and Ci-iooheteroalkynyl, wherein said Cuooheteroalkyl, Ci-iooheteroalkenyl and
- X-R 3 , X-R 4 or X-R 5 is a linking moiety selected from
- R" is methyl, methyloxy, hydroxy or halo
- n is 1. In another particular embodiment n, is 2. In another particular embodiment, each X is independently selected from -0-; -NH-; -NH-(CH2)-; -NH-(CH2)2-NH-;
- CH2-NH- wherein each of m and I are independently selected from 0, 1 , 2, 3, 4 and 5, and wherein Z is independently selected from -(CH 2 )i-i2-; -CH2-NH-CH2-; -CH2-CH 2 -NH-(CH 2 )i-3-; - CH2-CH2-N(-CH 3 )-CH2-CH 2 -; -CH 2 -NH-(CH 2 )2-3-NH-CH2-; -CH2-CH2-NH-(CH 2 )2 -NH-CH2-CH 2 -; - CH2-CH 2 -N(-CH 2 -CH2)2-N-CH 2 -CH2-; -CH2-NH-(CH 2 )2-NH-(CH 2 )2-NH-CH2-; -CH 2 -NH-(CH 2 ) 2 -NH- (CH 2 )2-NH-(CH 2 )2-NH-CH2-; -CH2-CH2-NH-CH2-NH-(CH2-NH-CH2-; -CH 2
- one of X-R 3 , X-R 4 or X-R 5 is a linking moiety selected from
- R" is methyl, methyloxy, hydroxy or halo
- Q is selected from H 2 N(CH 2 )30(CH2)20(CH2)3NH-, H 2 N(CH2)20(CH 2 )2NH-, H 2 N(CH 2 ) 3 0 (CH 2 ) 3 NH-, H 2 N(CH 2 )(CH 2 )1-12(CH2)NH-, ⁇ (CH ⁇ OiCH ⁇ OiCH ⁇ NH-, H 2 N(CH2) 2 NH(CH 2 )2NH- , H 2 N(CH2) 3 NH(CH 2 )i-4 NH-minister H 2 N(CH2)3 (CH 3 )(CH2)3NH-, ⁇ (CH ⁇ NHiCH ⁇ CH ⁇ NH-, H 2 N(CH2) 3 NH(CH2)2- (CH2)2NH(CH 2 )2NH -, H 2 N(CH 2 ) 3 N(CH2 CH 2 )NH(CH 2 ) 3 NH -,
- each of R 1a , R 1b and R 1c is methyl.
- each of R 1a , R 1b and R 1c is ethyl.
- one of X-R 3 , X-R 4 or X-R 5 can also be a linking moiety to which another substance or agent can be attached.
- Such linking moiety can be of any nature and length, as long as it allows another agent or substance or matrix to be coupled to it or to be associated with.
- the compounds of the invention are selected from the list of:
- each X is independently selected from -0-; -NH-; or -NH-(CH2)2-NH-;
- each X is independently selected from -NH-; or -NH-(CH2)2-NH-;
- each of R, R 3 , R 4 and R 5 is independently selected from dihydroxyphenyl or trihydroxyphenyl;
- each of R, R 3 , R 4 and R 5 is independently selected from a structure with formula (I);
- each Y is independently selected from hydrogen or methyl
- each X is independently selected from -NH-; or -NH-(CH2)2-NH-;
- each of R, R 3 , R 4 and R 5 is independently selected from a structure with formula (I)
- -k is 2; and (OY) is in the 3,4-position or the 4,5-position;
- -k is 3; and (OY) is in the 3,4,5-position or the 2,3,4-position;
- each X is independently selected from -NH-; or -NH-(CH 2 )2-NH-; - each of R, R 3 , R 4 and R 5 is independently selected from a structure with formula (I); -k is 3; and then (OY) is in the 3,4,5- position or the 2,3,4-position;and -each Y is hydrogen.
- a further particular embodiment is concerned with the use of the compounds for targeting HIV gp120
- n is independently selected from 1 and 2;
- - L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene,
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
- R 1a , R 1b and R 1c are independently selected from methyl; and ethyl;
- each of R, R 3 , R 4 and R 5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; dihydroxybenzyl; trihydrozybenzyl; dimethoxyphenyl; trimethoxyphenyl; dimethoxybenzyl;
- trimethoxybenzyl and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3 and wherein each Y is independently selected from hydrogen, methyl or benzyl; or
- R" is methyl, methyloxy, hydroxy or halo
- Q is selected from H 2 (CH 2 )30(CH2)20(CH2)3 H-, H 2 N(CH2)20(CH 2 )2NH-, H 2 N(CH 2 )30 (CH 2 ) 3 NH-, H 2 N(CH2)(CH 2 )i-i2(CH 2 )NH-, H 2 N(CH 2 )30(CH2)20(CH2)3NH-, H 2 N(CH2) 2 NH(CH 2 )2NH- , H 2 N(CH 2 )3NH(CH 2 )i4 NH-
- this further particular embodiment of the invention is concerned with the use of the compounds for targeting HIV gp120 wherein the compound is selected from the list of
- Another aspect of the invention is a pharmaceutical composition
- a pharmaceutical composition comprising a compound of the invention linked to an HIV inhibitor or antibody-recruiting therapeutic and a proces for preparing such a pharmaceutical composition 9 wherein a compound of the invention is intimately mixed with one or more pharmaceutical excipients or pharmaceutically acceptable carriers.
- the invention is further concerned with the above mentioned pharmaceutival compositions for the treatment of an HIV infection or the use of said composition for the manufacture of a medicament for the treatment of an HIV infection
- the invention also relates to a separation matrix for affinity chromatography of gp120, which matrix comprises the compounds of the invention.
- Another aspect of the present invention relates to a method for (a) binding or purifying (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus, or (3) gp120 comprising viruses or for (b) separating (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus or (3) gp120 comprising viruses, from other proteins, cells or viruses which do not comprise gp120, wherein said method comprises the use of a compound of the invention.
- the invention also relates to a method for quantifying, titrating or detecting gp120 of fragments thereof (to which the compounds bind).
- the invention further provides for the use of the compounds and for methods using the compounds for directing anti-viral drugs or other agents to gp120 comprising viruses or to gp120 comprising virus-infected cells.
- Yet another aspect relates to the use of the compounds of the invention for (a) binding or purifying (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus, or (3) gp120 comprising viruses or for (b) separating (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus or (3) gp120 comprising viruses, from other proteins, cells or viruses which do not comprise gp120, wherein said method comprises the use of a compound of the invention.
- the invention also relates to the use of the compounds of the invention for quantifying, titrating or detecting gp120 of fragments thereof (to which the compounds bind).
- virus comprising gp120 means a virus which comprises a gp120 gene and which exposes at least parts or fragments of the glycoprotein gp120 on its surface.
- gp120 comprising virus refers to HIV, the human immunodeficienccy virus which has the gp120 gene, env, and which exposes part of gp120 on the surface of the HIV envelope.
- association refers to a condition of proximity between a chemical entity, or portions thereof, and a target protein or peptide fragment thereof.
- the association may be non-covalent, wherein the juxta-position is energetically favoured by hydrogen bonding or van der Waals or electrostatic interactions, or alternatively it may be covalent.
- the number of carbon atoms represents the maximum number of carbon atoms generally optimally present in the substituent or linker; it is understood that where otherwise indicated in the present application, the number of carbon atoms represents the optimal maximum number of carbon atoms for that particular substituent or linker.
- LG means a chemical group which is susceptible to be displaced by a nucleophile or cleaved off or hydrolyzed in basic or acidic conditions.
- a leaving group is selected from a halogen atom (e.g., CI, Br, I) or a sulfonate (e.g., mesylate, tosylate, triflate).
- protecting group refers to a moiety of a compound that masks or alters the properties of a functional group or the properties of the compound as a whole.
- the chemical substructure of a protecting group varies widely.
- One function of a protecting group is to serve as intermediates in the synthesis of the parental drug substance.
- Chemical protecting groups and strategies for protection/deprotection are well known in the art. See: “Protective Groups in Organic Chemistry", Theodora W. Greene (John Wiley & Sons, Inc., New York, 1991. Protec' ' groups are often utilized to mask the reactivity of certain functional groups, to assist in the efficiency of desired chemical reactions, e.g. making and breaking chemical bonds in an ordered and planned fashion.
- Protection of functional groups of a compound alters other physical properties besides the reactivity of the protected functional group, such as the polarity, lipophilicity (hydrophobicity), and other properties which can be measured by common analytical tools.
- Chemically protected intermediates may themselves be biologically active or inactive.
- Protected compounds may also exhibit altered, and in some cases, optimized properties in vitro and in vivo, such as passage through cellular membranes and resistance to enzymatic degradation or sequestration. In this role, protected compounds with intended therapeutic effects may be referred to as prodrugs.
- Another function of a protecting group is to convert the parental drug into a prodrug, whereby the parental drug is released upon conversion of the prodrug in vivo.
- prodrugs may possess greater potency in vivo than the parental drug.
- Protecting groups are removed either in vitro, in the instance of chemical intermediates, or in vivo, in the case of prodrugs. With chemical intermediates, it is not particularly important that the resulting products after deprotection, e.g. alcohols, be physiologically acceptable, although in general it is more desirable if the products are pharmacologically innocuous.
- alkyl or "Ci-ie alkyl” as used herein means C1-C18 normal, secondary, or tertiary, linear or cyclic, branched or straight hydrocarbon with no site of unsaturation. Examples are methyl, ethyl, 1 -propyl (n-propyl), 2-propyl (iPr), 1 -butyl, 2-methyl-1 -propyl(i-Bu), 2-butyl (s- Bu), 2-dimethyl-2-propyl (t-Bu), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl- 2-butyl, 3-methyl-1 -butyl, 2-methyl-1 -butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl- 2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl,
- acyclic alkyl as used herein means C1-C18 normal, secondary, or tertiary, linear, branched or straight, hydrocarbon with no site of unsaturation. Examples are methyl, ethyl, 1 -propyl, 2-propyl (iPr), 1 -butyl, 2-methyl-1-propyl(i-Bu), 2-butyl (s-Bu), 2-methyl-2-propyl (t-Bu), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1 -butyl, 2- methyl-1 -butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pe 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-
- cycloalkyl or "C3-18 cycloalkyl” as used herein and unless otherwise stated means a saturated hydrocarbon monovalent radical having from 3 to 18 carbon atoms consisting of or comprising a C3-10 monocyclic or CMS polycyclic saturated hydrocarbon, such as for instance cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylethylene, methylcyclopropylene, cyclohexyl, cycloheptyl, cyclooctyl, cyclooctylmethylene, norbornyl, fenchyl,
- alkenyl or "CMsalkenyl” as used herein is C2-C18 normal, secondary or tertiary, linear or cyclic, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond.
- sites usually 1 to 3, preferably 1 of unsaturation, namely a carbon-carbon, sp2 double bond.
- the double bond may be in the cis or trans configuration.
- the term alkenyl refers to C1.12 hydrocarbons, yet more in particular to C1-6 hydro
- acyclic alkenyl refers to C 2 -Ci8 normal, secondary or tertiary, linear, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond.
- sites usually 1 to 3, preferably 1 of unsaturation, namely a carbon-carbon, sp2 double bond.
- the double bond may be in the cis or trans configuration.
- cycloalkenyl refers to a non-aromatic hydrocarbon radical having from 3 to 18 carbon atoms with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond and consisting of or comprising a C3-10 monocyclic or C7-18 polycyclic hydrocarbon. Examples include, but are not limited to:
- cyclopentenyl (-C5H7), cyclopentenylpropylene, methylcyclohexenylene and cyclohexenyl (-C6H9).
- the double bond may be in the cis or trans configuration.
- alkynyl or "CMsalkynyl” as used herein refers to C2-C18 normal, secondary, tertiary, linear or cyclic, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp triple bond.
- sites usually 1 to 3, preferably 1 of unsaturation, namely a carbon-carbon, sp triple bond. Examples include, but are not limited to: ethynyl (-C ⁇ CH), 3-ethyl-cyclohept-1 -ynylene, 4-cyclohept-1-yn-methylene and 1- propynyl (propargyl, -CH2C ⁇ CH).
- alkenyl refers to C1-12 hydrocarbons, yet more in particular to C1-6 hydrocarbons as further defined herein above.
- acyclic alkynyl refers to C2-C18 normal, secondary, tertiary, linear, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp triple bond. Examples include, but are not limited to: ethynyl (-C ⁇ CH) and 1-propynyl (propargyl, -CH2OCH).
- cycloalkynyl refers to a non-aromatic hydrocarbon radical having from 3 to 18 carbon atoms with at least one site (usually 1 to 3, preferably 1 ) of unsaturation, namely a carbon-carbon, sp triple bond and consisting of or comprising a C3-10 monocyclic or C7-18 polycyclic hydrocarbon. Examples include, but are not limited to: cyclohept-1- yne, 3-ethyl-cyclohept-1-ynylene, 4-cyclohept-1-yn-methylene and ethylene-cyclohept-1-yne.
- alkylene each refer to a saturated, branched or straight chain hydrocarbon radical of 1-18 carbon atoms (more in particular Ci-i2 or Ci-s carbon atoms), and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkane.
- Typical alkylene radicals include, but are not limited to: methylene (-CH 2 -), 1 ,2-ethyl (-CH2CH2-), 1 ,3-propyl (-CH2CH2CH2-), 1 ,4-butyl (- CH2CH2CH2CH2-), and the like.
- alkenylene each refer to a branched or straight chain hydrocarbon radical of 2-18 carbon atoms (more in particular C2-i2 or C2-6 carbon atoms) with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkene.
- alkynylene each refer to a branched or straight chain hydrocarbon radical of 2-18 carbon atoms (more in particular C2-i2 or C2-6 carbon atoms) with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp triple bond, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkyne.
- heteroalkyl refers to an acyclic alkyl wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
- heteroalkyl thus comprises -O-alkyl, -NH-alkyl, -N(alkyl)2, and -S-alkyl.
- heteroalkenyl refers to an acyclic alkenyl wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
- heteroalkenyl thus comprises -O-alkenyl, -NH-alkenyl, -N(alkenyl)2, -N(alkyl)(alkenyl), and -S-alkenyl.
- heteroalkynyl refers to an acyclic alkynyl wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
- heteroalkynyl tl comprises -0-alkynyl, -NH-alkynyl, -N(alkynyl)2, -N(alkyl)(alkynyl), -N(alkenyl)(alkynyl), and -S- alkynyl.
- heteroalkylene refers to an alkylene wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
- heteroalkenylene refers to an alkenylene wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
- heteroalkynylene refers to an alkynylene wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
- halogen means any atom selected from the group consisting of fluorine (F), chlorine (CI), bromine (Br) and iodine (I).
- Substituents optionally are designated with or without bonds. Regardless of bond indications, if a substituent is polyvalent (based on its position in the structure referred to), then any and all possible orientations of the substituent are intended.
- the present invention relates to a series of novel compounds which have been shown to have strong binding or association capacity to the glycoprotein gp120.
- the invention therefore relates to the new compounds, methods for their preparation, the use of the compounds for binding, purifying, titrating (quantifying), removing or separating gp120 and fragments thereof, cells infected with gp120 comprising viruses or viruses comprising gp120 and for targeting antiviral drugs or other substances or agents to gp120 (such as to gp120 comprising viruses).
- the present invention provides novel compounds which have a central benzene scaffold or a central carbohydrate scaffold, with a general structure according to the formulas (A), (B), (C), (D) or (E),
- n is independently selected from 1 and 2;
- - L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene,
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
- heteroalkynylene and (1 ,4-piperazinylene);
- each of R 1a , R 1b and R 1c is independently selected from methyl; and ethyl;
- each of R, R 3 , R 4 and R 5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3; or
- - one of X-R 3 , X-R 4 or X-R 5 is a linking moiety to which another substance or agent can be attached.
- the compounds of the invention can have a central benzene scaffold or a central carbohydrate scaffold and a variation of compounds with these central scaffolds are envisaged in this invention.
- Compounds of formula AA containing 2,4,6-triethyl(methyl)benzene as central scaffold are envisaged with this invention.
- di-and trihydroxy benzoyl I and caffeoyl II moieties are introduced and in addition, some non aromatic polyhydroxy residues structurally related to I such as shikimyl III or quinyl IV are introduced. These residues are connected to the central scaffold by ester or amide bonds. verview 1 : compounds of formula AA
- R2 R3 F3 ⁇ 4
- D-Glucose As monosaccharides D-Glucose, D-Galactose, D-Allose, D-Mannose, D-Xylose, D-Arabinose, D- Ribose and D-Lyxose are envisaged. Disaccharides composed of glucose: cellobiose, maltose, gentibiose and trehalose are also contemplated. In addition, disaccharides made up of glucose and other sugars such as lactose (galactose-glucose) and sucrose (glucose-fructose) are envisaged. In both series of compounds, aromatic moieties (I and II) and non aromatic polyhydroxy residues (III and IV) are connected to the central scaffolds by ester bonds.
- Compounds can furthermore be linked (i.e. directly or through a spacer molecule) to a therapeutic drug or to a dye, a fluorescent molecule, a diagnostic enzyme, or a radiolabeled entity to enable identification of gp120 containing viruses or virus-infected (gp120 expressing) cells or to target therapeutically gp120 containing viruses or virus-infected (gp120 expressing) cells.
- a spacer molecule i.e. directly or through a spacer molecule
- the compounds of the invention can thus be used to bind, purify, titrate, quantify, remove or separate gp120 or fragments thereof, cells infected with gp120 comprising viruses or viruses comprising gp120 and can be used for targeting antiviral drugs or other substances or agents to gp120 (such as to gp120 comprising viruses).
- compositions of the invention relate to a pharmaceutical composition
- a pharmaceutical composition comprising a compound of the invention as a carrier for targeting antiviral drugs or other substances or agents to gp120 (such as to gp120 comprising viruses or cells).
- the compositions of the invention may be administered to the mammal (including a human) to be treated by any means well known in the art, i.e. orally, intranasally, subcutaneously, intramuscularly, intradermal ⁇ , intravenously, intra-arterially, parenterally or by catheterization.
- the invention also relates to a process of preparing a pharmaceutical composition of the invention wherein the compounds of the invention, carrying the one or more therapeutical antiviral drugs or other substances or agents and the optionally one or more pharmaceutical excipients or pharmaceutically acceptable carriers are intimately mixed.
- the invention further relates to the use of a composition
- a composition comprising (a) one or more derivatives of formula (A), (B), (C), (D) or (E) as carrier for targeting one or more viral inhibitors to provide a synergistic effect against a viral infection, preferably a HIV/AIDS viral infection.
- the viral inhibitors may be
- Carbohydrate-binding agents i.e. pradimicins, lectins, 2G12
- Anionic compounds i.e. PRO-2000, PVAS, PAVAS
- the viral inhibitors or other substances or agents may be antibody-recruiting therapeutics such as but not limited to:
- the bifunctional molecule that is obtained by coupling the compounds of the inventions to the above mentioned antibody-recruiting therapeutics can recruit antibodies such as but not limited to:
- the invention also relates to the compositions of the invention being used for inhibition of the replication of HIV strains.
- the invention relates to the compositions of the invention being useful as agents having biological activity (preferably antiviral activity) or as diagnostic agents.
- biological activity preferably antiviral activity
- diagnostic agents any of the uses mentioned with respect to the present invention may be restricted to a non-medical use, a non- therapeutic use, a non-diagnostic use, or exclusively an in vitro use, or a use related to cells remote from an animal.
- the invention thus relates to a method for the treatment of viral infections in humans and other mammals wherein a therapeutically effective amount of the composition is administered to a human or a mammal in need of said treatment.
- the therapeutically effective amount especially for the treatment of viral infections in humans and other mammals, is a HIV protein inhibiting amount. More particularly, it is a HIV replication inhibiting amount.
- the compounds of the invention can be linked to or associated with (absorbed on or into) a polymer or a matrix used in column chromatography, such as in affinity chromatography in order to create a new matrix for column chromatography for binding, separating or purifying gp120 or fragments thereof.
- the compounds of the invention is to be used in a form immobilised to a solid support, e.g. as a ligand in affinity chromatography for gp120, wherein the compounds or a linker attached thereto comprises terminating functionalities useful for such immobilisation.
- Another embodiment relates to a separation matrix for use in affinity chromatography, wherein the ligands of the separation mix comprise at least one compound as defined above.
- the ligands have been coupled to a support via linkers.
- the present matrix can e.g. be in the form of separate particles, preferably porous and essentially spherical particles; a monolith; or a membrane.
- a system suitable for affinity chromatography which is comprised of a separation matrix as defined above packed in a column.
- the column may be of a size suitable for analytical scale or for large scale chromatography.
- the support is a natural polymer, such as agarose, alginate, carrageenan, gelatine etc.
- agarose alginate
- carrageenan a natural polymer
- Such natural polymers are known to form physically cross-linked networks spontaneously on cooling or on addition of divalent metal ions, and chemical cross-linkers can be added if desired.
- This kind of supports is easily prepared according to standard methods, such as inverse suspension gelation (S Hjerten, Biochim Biophys Acta 79(2), 393-398 (1964)).
- the support is comprised of cross-linked synthetic polymers, such as styrene or styrene derivatives, divinylbenzene, acrylamides, acrylate esters, methacrylate esters, vinyl esters, vinyl amides etc.
- polymers are also easily produced according to standard methods, see e.g. "Styrene based polymer supports developed by suspension polymerization” (R Arshady: Chimica e L'lndustria 70(9), 70-75 (1988)).
- the support material can in principle be any material that allows the covalent coupling of the gp120 binding compounds of the invention, such as the above-discussed polymers, inorganic materials, such as silica, ceramics etc.
- the support has hydrophilic surfaces, and if porous, the surfaces of the pores are also hydrophilic. This is advantageous in order to avoid or at least reduce any non-specific protein interactions. It is also advantageous if the surfaces have a high density of groups available for coupling of ligands.
- Such coupling groups are commonly hydroxyl groups, but may also be ally, groups i.e. double bonds available for grafting, amines, thioles, epoxides and the like.
- the support material has undesirable surface properties, it is possible to coat it with a hydrophilic polyhydroxy- functional material before coupling the ligand.
- the techniques and considerations for coupli affinity ligands to a suitable support to prepare a separation matrix are well known in this field, see e.g. WO 98/33572 for a detailed review of coupling chemistry as well as suitable linking molecules.
- the compounds can be used in a method of isolating or separating gp120 or fragments thereof, from other components, for example in a liquid, wherein a compound or a separation matrix as defined above is used.
- the present method is affinity chromatography, which is a widely used and well-known separation technique.
- a solution comprising gp120 or binding fragments thereof is passed over a separation matrix under conditions allowing adsorption of gp120 or fragments thereof to the compounds of the present invention present on said matrix.
- Such conditions are controlled e.g. by pH and/or salt concentration i.e. ionic strength in the solution. Care should be taken not to exceed the capacity of the matrix, i.e.
- the flow should be sufficiently slow to allow a satisfactory adsorption.
- other components of the solution will pass through in principle unimpeded.
- the matrix is then washed, e.g. with an aqueous solution, in order to remove retained and/or loosely bound substances.
- a second solution denoted an eluent is passed over the matrix under conditions that provide desorption i.e. release of gp120 or fragments thereof. Such conditions are commonly provided by a change of the pH, the salt concentration i.e. ionic strength, hydrophobicity etc.
- Various elution schemes are known, such as gradient elution and step-wise elution. Elution can also be provided by a second solution comprising a competitive substance, which will replace the gp120 or fragments thereof on the matrix.
- the compounds of the invention is used in an assay for detection of gp120, fragments thereof or gp120 comprising cells.
- the compound is preferably labelled with a suitable detectable label as conventionally used, such as a fluorescent label, a luminescent label, a chemiluminiscent label, an enzyme label, a radioactive label, an absorbance label etc.
- the compounds of the invention can be linked to anti-HIV entry inhibitors in order to target to the anti-HIV drugs to the site of interaction and to retain the anti-HIV drugs for a longer period at their site of interaction.
- the compounds of the invention according to the formulae of the application can be prepared while using a series of chemical reactions known to those skilled in the art, altogether making up the process for preparing said compounds and exemplified further.
- the proce described further are only meant as examples and by no means are meant to limit the scope of the present invention.
- the gp120 binding compounds can be linked (whether or not via a linker) to a therapeutic drug, on a column polymer, to a dye, a fluorescent molecule, a diagnostic enzyme, or a radiolabeled entity to enable identification of gp120 containing viruses or virus-infected.
- a linker moiety can be performed as shown in scheme 7.
- the therapeutic drug, polymer, dye, fluorescent molecule, diagnostic enzyme, or radiolabeled entity, amongh others can be coupled to the linker through reactions known in the art.
- EXAMPLE 1 GENERAL SYNTHETIC METHODS AND MATERIALS FOR THE PREPARATION OF THE COMPOUNDS WITH A CENTRAL BENZENE SCAFFOLD
- Acetonitrile was used as mobile phase A with 0.08% of formic acid, and water was used as mobile phase B with n ⁇ ⁇ ' of formic acid at a flow rate of 1 ml/min.
- Two different methods were used, one on a XBridge Cie (2.1 x 100 mm, 3.5 ⁇ ) column and A:B, 5-80% of A that will be noted as f/?pg and the other on a SunFire Cie (4.6 x 50 mm, 3.5 ⁇ ) column with a A:B, 0-100% of A, that will be noted as t ⁇ s). Melting points were measured on a Reichert-Jung Kofler. Optical rotations were measured on a Perkin-Elmer 241 MC polarimeter with a cell of path length 1 dm.
- JP 08143525 A General procedure A for the reaction of trisamine 1 with benzoic acids (in accordance with Scheme 1).
- Benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (BOP) (3.3 eq) was added to a solution of the corresponding benzoic acid (3.3 eq) in dry dichloromethane (5 mL).
- trisamine 1 50 mg, 1 eq
- triethylamine 3.3 eq
- EXAMPLE 6 Synthesis of 1,3,5-Tris(3,4-dimethoxyphenylaminomethyl)-2,4,6- triethylbenzene (15, AL-125). To a solution of 11 (100 mg, 0.22 mmol) and the commercially available 3,4-dimethoxyaniline 12 (139.4 mg, 0.91 mmoles) in acetonitrile (6 mL) was added dry potassium carbonate (125.8 mg, 0.91 mmol) and the mixture was refluxed overnight. After evaporation of the solvent, the residue was dissolved in ethyl acetate (15 mL) and washed with water (3 x15 mL).
- EXAMPLE 9 Synthesis of 1,3,5-Tris(3,4,5-trimethoxybenzylaminomethyl)-2,4,6- triethylbenzene (18, AL 177).
- a solution of 11 (167 mg, 0.37 mmol) and the commercially available 3,4,5-trimethoxybenzylamine 14 (262 mg, 1.3 mmol) was treated according to the general procedure B.
- the residue was purified by CCTLC (dichloromethane/methanol/NH 4 OH, 9:1 :0.2) to give 0.19 g (65%) of 18, AL 177 as a white solid: mp 100-102 °C.
- HPLC: fo w 2.78.
- Benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (48 mg, 0.11 mmol, 1.2 equiv.) was added to a solution of 2,3,4-tris(benzyloxy)benzoic acid 3 (40 mg, 0.09 mmol) and triethylamine (15 ⁇ , 0.11 mmol, 1.2 equiv.) in dry dichloromethane (3 ml_). After 1.5 h, the corresponding 1 ,2-diaminoetane (5.9 ⁇ , 0.09 mmol) was added. The mixture was stirred at room temperature overnight; during this time a white solid was formed.
- EXAMPLE 12 Synthesis of 1 ,3,5-Tris(2,3,4-tribenzyloxybenzamidoethylaminomethyl)-2,4,6- triethylbenzene (21, ECM-VI-27). A solution of 11 (50 mg, 0.11 mmol) and /V-(2-aminoethyl)- 2,3,4-tris(benzyloxy)benzamide 20 (231 mg, 0.47 mmol) was treated with triethylamine (0.065 mL, 0.47 mmol) according to the general procedure B.
- EXAMPLE 13 Synthesis of 1,3,5-Tris(2,3,4-trihydroxybenzamidoethylaminomethyl)-2,4,6- triethylbenzene (22, AL-184). Following the general procedure, the benzyl derivative 21, ECM- VI-27 (92 mg, 0.06 mmol) gave 45 mg (96 %) of 22, AL-184 as a white solid: mp 164-166 °C.
- EXAMPLE 14 GENERAL SYNTHESIS METHODS AND MATERIALS FOR SYNTHESIS OF THE COMPOUNDS WITH A CENTRAL CARBOHYDRATE SCAFFOLD ⁇ protected mono- and disaccharides.
- EXAMPLE 15 Synthesis of or- and j3-D-Mannopyranose pentakis[2,3,4- tris(phenylmethoxy)benzoate] (35 and 36 PCB248). Following the general procedure D- mannose (50 mg, 0.28 mmol) was treated with 3 (713 mg, 1.62 mmoles, 6 eq), DCC (33,4 mg, 1.62 mmol, 6 eq) and DMAP (198 mg, 1.62 mmol, 6 eq) to give a residue that was purified by column chromatography on silica gel using a mixture of toluene/ethyl acetate (96:4).
- EXAMPLE 18 Synthesis of 0-D-Maltose octakis[3,4,5-tris(phenylmethoxy)benzoate] (39 AL34). Following the general procedure monohydrated Maltose (50 mg, 0.14 mmol) was treated with 2 10 (607 mg, 1.38 mmoles, 10 eq), DCC (284 mg, 1.38 mmol, 10 eq) and DMAP (169 mg, 1.38 mmol, 10 eq) to give a residue that was purified by column chromatography on silica gel using a mixture of dichloromethane/toluene/ethyl acetate 75:25:1 to give 440 mg (86%) of 39 AL34 as a white solid.
- EXAMPLE 19 Synthesis of /J-D-Maltose octakis(3,4,5-trihydroxybenzoate) (40, AL175).
- EXAMPLE 20 Synthesis of ⁇ , ⁇ -Trehalose octakis[3,4,5-tris(phenylmethoxy)benzoate] (41, AL35). Following the general procedure dehydrated ⁇ , ⁇ -Trehalose (50 mg, 0.14 mmol) was treated with 2 (581 mg, 1.32 mmoles, 10 eq), DCC (272.3 mg, 1.32 mmol, 10 eq) and DMAP (161.26 mg, 1.32 mmol, 10 eq) to give a residue that was purified by column chromatography on silica gel using a mixture of dichloromethane/toluene/ethyl acetate 75:25:1 to give 398 mg (81%) of 41 AL35 as a white solid.
- Recombinant gp120 protein from the HIV-1 NIB strain (ImmunoDiagnostics Inc., Woburn, MA) (produced in cell cultures of Chinese hamster ovaries) was covalently immobilized on the carboxymethylated dextran matrix of a CM5 sensor chip in 10 mM sodium acetate, pH 4.0 using standard amine coupling chemistry up to a final density of 1770 RUs (15 fmol of gp120).
- a reference flow cell was used as a control for non-specific binding and refractive index changes. All interaction studies were performed at 25 °C on a Biacore T100 instrument (GE Healthcare, Uppsala, Sweden).
- the compounds were diluted in HBS-P (10 mM HEPES, 150 mM NaCI and 0,05% surfactant P20; pH 7.4) supplemented with 5% dimethyl sulfoxide (DMSO, Merck) and 10 mM Ca 2+ at a concentration of 50 ⁇ . Samples were injected for 3 minutes at a flow rate of 30 ⁇ /min followed by a dissociation phase of 5 minutes. A DMSO concentration series was included to eliminate the contribution of DMSO to the measured response. The CM5 sensor chip surface was regenerated with an injection of 50 mM NaOH for 5 seconds at a flow rate of ⁇ /min. The compounds that showed significant binding to gp120 are shown in table 1. The affinity of the compounds is evaluated through the response units obtained from the binding of the compounds to the chip both at the end of the injection phase and after 50s of dissociation.
- DMSO dimethyl sulfoxide
- Table 1 Response units (RUs) of the binding of the synthesized compounds to gp120 on a CM5 sensor chip at the end of the injection phase and after 50 sec of dissociation.
- N-Fmoc-N'- succinyl-4,7,10-trioxa-1 ,13-tridecanediamine 104 (232 mg, 0.43 mmol) (Reina, J.J., et al. Bioconjugate Chem. 2007, 18, 963-969) and EtjN (60 ⁇ , 0.43 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 1h. Dichloromethane (50 mL) was added and the reaction mixture was washed with brine (20 mL).
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Abstract
The present invention provides for novel benzene compounds and saccharide compounds and for the use of said compounds for binding, titration (quantification), removing, purifying or separating the glycoprotein gp120, gp120 comprising viruses or cells infected with gp120 comprising viruses. The invention also provides for a method for the detection, binding, titration (quantification), removal, purification or separation of (or directing therapeutic or other agents to) gp120, gp120 comprising viruses or cells infected with gp120 comprising viruses. The invention further provides for the use of the compounds and for methods using the compounds for directing anti -viral drugs or other agents to gp120 comprising viruses or to gp120 comprising virus - infected cells. The present invention also provides processes for the preparation of said novel compounds.
Description
GP120 -BINDING BENZENE COMPOUNDS AND SACCHARIDE COMPOUNDS
FIELD OF THE INVENTION
The present invention provides for novel compounds and for the use of said compounds for binding, titration (quantification), removing, purifying or separating the glycoprotein gp120, gp120 comprising viruses or cells infected with gp120 comprising viruses. The invention also provides for a method for the detection, binding, titration (quantification), removal, purification or separation of gp120, gp120 comprising viruses or cells infected with gp120 comprising viruses. The invention thus also relates to a separation matrix for affinity chromatography of gp120, which matrix comprises the compounds of the invention. The invention further provides for the use of the compounds and for methods using the compounds for directing anti-viral drugs or other agents to gp120 comprising viruses and gp120 comprising virus-infected cells.
The present invention also provides processes for the preparation of said novel compounds.
BACKGROUND OF THE INVENTION
Gp120 is a glycoprotein exposed on the surface of the HIV envelope. The 120 in its name comes from its molecular weight of 120 kilodaltons. gp120 is essential for virus entry into cells as it plays a vital role in seeking out specific cell surface receptors for entry. The gp120 gene, env, is around 1.5 kb long and codes for around 500 amino acids. Three gp120s, bound as heterotrimers to a transmembrane glycoprotein, gp41, are thought to combine in a trimer to form the envelope spike, which is involved in virus-cell attachment.
The Human Immunodeficiency Virus (HIV) can mutate frequently to stay ahead of the immune system. There is however a highly conserved region in the virus genome near its receptor binding site. The glycoprotein gp120 is anchored to the viral membrane, or envelope, via non-covalent bonds with the transmembrane glycoprotein, gp41. It is involved in entry into cells by binding to CD4 receptors, particularly helper T-cells and macrophages. Binding to CD4 is mainly electrostatic although there are van der Waals interactions and hydrogen bonds.
Since gp120 plays a vital role in the ability of HIV-1 to enter CD4+ cells, its evolution is of particular interest. Many neutralizing antibodies bind to sites located in variable regions of gp120. The diversity of env has been shown to increase by 1-2% per year in HIV-1 group M and the variable units are notable for rapid changes in amino acid sequence length. Increases in gp120
variability result in significantly elevated levels of viral replication, indicating an increase in viral fitness in individuals infected by diverse HIV-1 variants. Further studies have shown that variability in potential N-linked glycosylation sites (PNGSs) also result in increased viral fitness. PNGSs allow for the binding of long-chain carbohydrates to the high variability regions of gp120, so it is possible that the number of PNGSs in env might affect the fitness of the virus by providing more or less sensitivity to neutralizing antibodies. The presence of large carbohydrate chains extending from gp120 might obscure possible antibody binding sites. The relationship between gp120 and neutralizing antibodies is an example of Red Queen evolutionary dynamics. Continuing evolutionary adaptation is required for the viral envelope protein to maintain fitness relative to the continuing evolutionary adaptations of the host immune neutralizing antibodies, and vice-versa, forming a coevolving system.
Since CD4 receptor binding is the most obvious step in HIV infection, gp120 was among the first targets of HIV vaccine research. Efforts to develop HIV vaccines targeting gp120, however, have been hampered by the chemical and structural properties of gp120, which make it difficult for antibodies to bind to it. gp120 can also easily be shed from the surface of the virus and captured by T cells due to its loose binding with gp41. Also research is being performed to identify a new vaccine based on antibodies that hydrolyze or cleave a part of the gp120 protein, rendering it incapable of binding to lymphocytes. This binding is the first step in the process of HIV infection. The protein gp120 is necessary during the initial binding of HIV to its target cell. Consequently, anything which binds to gp120's target can block gp120 from binding to a cell by being physically in the way. Many of these are toxic to the immune system, such as the anti-CD4 monoclonal antibody OKT4. Also low molecular weight compounds have been developed that bind to the same CD4 receptor that gp120 binds to.
However, low molecular weight compounds that bind to gp120 in order to block its function in the HIV infection cycle are rather rare. Furthermore, gp120 is a large glycoprotein for which the use of a gp120 binder could be helpful in the purification of gp120. Gp120 could also be targeted in order to identify, bind, purify or separate gp120 comprising viruses or cells infected with gp120 comprising viruses, such as HIV. Finally, compounds can be used for directing anti-viral drugs or other agents to gp120 comprising viruses.
SUMMARY OF THE INVENTION
The present invention relates to novel gp120 binding compounds. The compounds have been found to bind to gp120 and can therefore be used for binding, titration (quantification, removal, purification or separation of gp120. The compounds can also be used to titrate (quantify), remove, detect, bind to, purify or separate gp120 comprising viruses or cells infected with gp120 comprising viruses, such as HIV. The compounds can furthermore be used for directing or targeting anti-viral drugs or other agents to gp120 comprising viruses or to gp120 comprising virus-infected cells.
The first aspect of the present invention provides novel compounds which have a central benzene scaffold or a central carbohydrate scaffold, with a general structure according to the formulas (A), (B), (C), (D) or (E),
(B)
(D) (E)
wherein,
- each n is independently selected from 1 and 2;
- L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene,
heteroalkenylene, and heteroalkynylene can be unsubstituted or substituted with =0, =S; OH; - SH; and NH2;
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
— N—
heteroalkynylene; and (1 ,4-piperazinylene);
- each of R1a, R1b and R1c is independently selected from methyl; and ethyl;
- each of R, R3, R4 and R5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; dihydroxybenzyl; trihydrozybenzyl; dimethoxyphenyl; trimethoxyphenyl; dimethoxybenzyl;
trimethoxybenzyl, and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3 and wherein each Y is independently selected from hydrogen, methyl or benzyl; or
- one of X-R3, X-R4 or X-R5 is a linking moiety selected from Ci-iooheteroalkyl; Ci-iooheteroalkenyl; and Ci-iooheteroalkynyl, wherein said Cuooheteroalkyl, Ci-iooheteroalkenyl and
Ci-iooheteroalkynyl can be unsubstituted or substituted with one or more =0, =S; OH; -SH; and NH2; or
- one of X-R3, X-R4 or X-R5 is a linking moiety selected from
wherein Q is selected from H2N(CH2)30(CH2)20(CH2)3NH-, H2N(CH2)20(CH2)2NH-, H2N(CH2)30 (CH2)3NH-, H2N(CH2)(CH2)i-i2(CH2)NH-, H2N(CH2)30(CH2)20(CH2)3NH-, H2N(CH2)2NH(CH2)2NH- , H2N(CH2)3NH(CH2)i NH-„ H2N(CH2)3N(CH3)(CH2)3NH-, H2N(CH2)2NH(CH2)2-3(CH2)2NH-, H2N(CH2)3NH(CH2)2 (CH2)2NH(CH2)2NH -, H2N(CH2)3N(CH2 CH2)NH(CH2)3NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)3NH(CH2)CH=CH(CH2)NH(CH2)3NH -;
with the proviso that said compound is not
- 1 ,3,5-Tris(2,3-dimethoxybenzamidomethyl)-2,4,6-triethylbenzene (9, PCB311);
- 1 ,3,5-Tris(2,3-dihydroxybenzamidomethyl)-2,4,6-triethylbenzene (10, AL180);
- or- or β-D-Glucopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (23, AL-18);
- a- or β-D-Glucopyranose pentakis[3,5-bis(phenylmethoxy)benzoate];
- a- or β-D-Glucopyranose pentakis[3,4-bis(phenylmethoxy)benzoate];
- σ or β-D-Glucitol 1,5-anhydro-pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or -D-Galactopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (25, AL-29 and AL-30);
- a- or j8-D-Allopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or -D-Mannopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate](29, 30 and 31);
- a- or j6-D-Xylopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or /?-L-Arabinopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- or- or -D-Ribopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate] (27, AL36);
- a- or jS-D-Luxopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or jS-D-Glucopyranose pentakis[3,5-dihydroxybenzoate];
- a- or jS-D-Glucopyranose pentakis[3,4-dihydroxybenzoate];
- D-Galactopyranose pentakis[3,4-dihydroxybenzoate];
- D-Arabinopyranose tetrakis[3,4-dihydroxybenzoate];
- D-Mannopyranose pentakis[3,4-dihydroxybenzoate];
- a or jS-D-Glucitol 1 ,5-anhydro-pentakis[3,4,5-trihydroxybenzoate];
- a- or jS-D-Glucopyranose pentakis[3,4,5-trihydroxybenzoate] (24, AL-19);
- a- or -D-Galactopyranose pentakis[3,4,5-trihydroxybenzoate] (26, AL-33);
- a- or jS-D-Allopyranose pentakis[3,4,5-trihydroxybenzoate];
- or- or j8-D-Mannopyranose pentakis[3,4,5-trihydroxybenzoate](32, AL51a and 33, AL51b);
- a- or j8-D-Xylopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or jS-L-Arabinopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or j8-D-Ribopyranose tetrakis[3,4,5-trihydroxybenzoate] (28, AL50);
- a- or j8-D-Luxopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or j8-D-Glucopyranose pentakis[3,4,5-trimethoxybenzoate];
- σ- or jS-D-Mannopyranose pentakis[3,4,5-trimethoxybenzoate]
-j8-D-Glucopyranose1-[4,5-dihydroxy-2-(phenylmethoxy)benzoate]2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate];
- β -D-Glucopyranose, 1-[3,4-dihydroxy-5-(phenylmethoxy)benzoate] 2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate].
In a particular embodiment, n is 1. In another particular embodiment n, is 2. In another particular embodiment, each X is independently selected from -0-; -NH-; -NH-(CH2)-; -NH-(CH2)2-NH-;
0-(CH2)i-i8-0-; -0-(CH2)2-0-(CH2)2-0-; -NH-(CH2)3-0-(CH2)2-0-(CH -0-(CH2)3-NH-
CH2OVCH2-CH2-NH-; -NH-CH2-CH2-CH2-(0-CH2-CH2)i-CH2-NH-;
; -NH-CH2-Z-
CH2-NH-, wherein each of m and I are independently selected from 0, 1 , 2, 3, 4 and 5, and
wherein Z is independently selected from -(CH2)i-i2-; -CH2-NH-CH2-; -CH2-CH2-NH-(CH2)i-3-; - CH2-CH2-N(-CH3)-CH2-CH2-; -CH2-NH-(CH2)2-3-NH-CH2-; -CH2-CH2-NH-(CH2)2 -NH-CH2-CH2-; - CH2-CH2-N(-CH2-CH2)2-N-CH2-CH2-; -CH2-NH-(CH2)2-NH-(CH2)2-NH-CH2-; -CH2-NH-(CH2)2-NH- (CH2)2-NH-(CH2)2-NH-CH2-; -CH2-CH2-NH-CH2-CH=CH-CH2-NH-(CH2)2-.
In another particular embodiment one of X-R3, X-R4 or X-R5 is a linking moiety selected from
wherein R" is methyl, methyloxy, hydroxy or halo; and
wherein Q is selected from H2N(CH2)30(CH2)20(CH2)3NH-, H2N(CH2)20(CH2)2NH-, H2N(CH2)30 (CH2)3NH-, H2N(CH2)(CH2)1-12(CH2)NH-, ^(CH^OiCH^OiCH^NH-, H2N(CH2)2NH(CH2)2NH- , H2N(CH2)3NH(CH2)i-4 NH-„ H2N(CH2)3 (CH3)(CH2)3NH-, ^(CH^NHiCH^CH^NH-, H2N(CH2)3NH(CH2)2- (CH2)2NH(CH2)2NH -, H2N(CH2)3N(CH2 CH2)NH(CH2)3NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)3NH(CH2)CH=CH(CH2)NH(CH2)3NH -.
In another particular embodiment, each of R1a, R1b and R1c is methyl.
In another particular embodiment, each of R1a, R1b and R1c is ethyl.
In a particular embodiment, one of X-R3, X-R4 or X-R5 can also be a linking moiety to which another substance or agent can be attached. Such linking moiety can be of any nature and length, as long as it allows another agent or substance or matrix to be coupled to it or to be associated with. In yet another more particular embodiment, the linking moiety is selected from Ci-iooheteroalkyl; Ci-iooheteroalkenyl; and Ci-iooheteroalkynyl, wherein said Ci-iooheteroalkyl, Ci. looheteroalkenyl and Ci-iooheteroalkynyl can be unsubstituted or substituted with one or more =0, =S; OH; -SH; and NH2.
In yet another embodiment, the compounds of the invention are selected from the list of:
- 1 ,3,5-Tris(3,4,5-tribenzyloxybenzamidomethyl)-2,4,6-triethylbenzene (5, PCB245);
- 1 ,3,5-Tris(3,4,5-trihydroxybenzamidomethyl)-2,4,6-triethylbenzene (6, AL 113);
- 1 ,3,5-Tris(2,3,4-tribenzyloxybenzamidomethyl)-2,4,6-triethylbenzene (7, AL 168);
- 1 ,3,5-Tris(2,3,4-trihydroxybenzamidomethy!)-2,4,6-triethyibenzene (8, AL 170);
- 1 ,3,5-Tris(3,4-dimethoxyphenylaminomethyl)-2,4,6-triethylbenzene (15, AL-125);
- 1 ,3,5-Tris(3,4-dihydroxyphenylaminomethyl)-2,4,6-triethylbenzene (16, AL-172);
- 1 ,3,5-Tris(3,4,5-trimethoxyphenylaminomethyl)-2,4,6-triethylbenzene (17, AL 126);
- 1 ,3,5-Tris(3,4,5-trimethoxybenzylaminomethyl)-2,4,6-triethylbenzene (18, AL 177);
- 1 ,3,5-Tris(3,4,5-trihydroxybenzylaminomethyl)-2,4,6-triethylbenzene (19, AL 178);
- 1 ,3,5-Tris(2,3,4-tribenzyloxybenzamidoethylaminomethyl)-2,4,6-triethylbenzene (21 , ECM-VI- 27);
- 1 ,3,5-Tris(2,3,4-trihydroxybenzamidoethylaminomethyl)-2,4,6-triethylbenzene (22, AL-184);
- a- or jS-D-Mannopyranose pentakis[2,3,4-tris(phenylmethoxy)benzoate] (35 and 36 PCB248);
- a- D-Mannopyranose pentakis(2,3,4-trihydroxybenzoate) (37, AL-173);
- β- D-Mannopyranose pentakis(2,3,4-trihydroxybenzoate) (38, AL-174);
- β- D-Maltose octakis[3,4,5-tris(phenylmethoxy)benzoate] (39, AL34);
- β- D-Maltose octakis(3,4,5-trihydroxybenzoate) (40, AL175);
- a, a-Trehalose octakis[3,4,5-tris(phenylmethoxy)benzoate] (41 , AL35);
- a, a-Trehalose octakis (3,4,5-trihydroxybenzoate) 42 (AL166); and
- 1-A/-[ (13 - amino-4',7',10'-trioxatridecyl)succinamoyl]aminomethyl-3,5-Bis(2,3,4- trihydroxybenzoylaminomethyl)-2,4,6-triethylbenzene.
Interesting compounds are the compounds of formula (A) or (B).
Other interesting compounds are the compounds of formula (C), (D), or (E).
More interesting compounds encompass those particular embodiments and those interesting compounds wherein one or more of the following restriction apply:
- each X is independently selected from -0-; -NH-; or -NH-(CH2)2-NH-;
- each X is independently selected from -NH-; or -NH-(CH2)2-NH-;
- each of R, R3, R4 and R5 is independently selected from dihydroxyphenyl or trihydroxyphenyl;
- each of R, R3, R4 and R5 is independently selected from a structure with formula (I);
-k is 2; and then (OY) is in the 3,4-position or the 3,5-position;
-k is 2; and then (OY) is in the 3,4-position:
-k is 3; and then (OY) is in the 3,4,5-position or the 2,3,4-position;
-k is 3; and then (OY) is in the 2,3,4-position;
-k is 3; and then (OY) is in the 3,4,5-position;
-each Y is independently selected from hydrogen or methyl;
Even more interesting compounds encompass those particular embodiments, those interesting compounds and those more interesting compounds wherein one or more of the following restriction apply:
- each X is independently selected from -NH-; or -NH-(CH2)2-NH-;
- each of R, R3, R4 and R5 is independently selected from a structure with formula (I)
-k is 2; and (OY) is in the 3,4-position or the 4,5-position;
-k is 3; and (OY) is in the 3,4,5-position or the 2,3,4-position;
-each Y is hydrogen.
Most interesting compounds are those compounds of the invention wherein each X is independently selected from -NH-; or -NH-(CH2)2-NH-; - each of R, R3, R4 and R5 is independently selected from a structure with formula (I); -k is 3; and then (OY) is in the 3,4,5- position or the 2,3,4-position;and -each Y is hydrogen.
A further particular embodiment is concerned with the use of the compounds for targeting HIV gp120
wherein the compound has a structure according to formula (A), (B), (C), (D), or (E)
(C) (D) (E) wherein,
- each n is independently selected from 1 and 2;
- L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene,
heteroalkenylene, and heteroalkynylene can be unsubstituted or substituted with =0, =S; OH; SH; and NH2;
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
heteroalkynylene; and 1 ,4-piperazinylene;
- each of R1a, R1b and R1c is independently selected from methyl; and ethyl;
- each of R, R3, R4 and R5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; dihydroxybenzyl; trihydrozybenzyl; dimethoxyphenyl; trimethoxyphenyl; dimethoxybenzyl;
trimethoxybenzyl, and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3 and wherein each Y is independently selected from hydrogen, methyl or benzyl; or
- one of X-R3, X-R4 or X-R5 is a linking moiety selected from Ci-iooheteroalkyl; Ci-iooheteroalkenyl; and Ci-iooheteroalkynyl, wherein said Ci-iooheteroalkyl, Ci-iooheteroalkenyl and Ci- looheteroalkynyl can be unsubstituted or substituted with one or more =0, =S; OH; -SH; and NH2; or
wherein Q is selected from H2 (CH2)30(CH2)20(CH2)3 H-, H2N(CH2)20(CH2)2NH-, H2N(CH2)30 (CH2)3NH-, H2N(CH2)(CH2)i-i2(CH2)NH-, H2N(CH2)30(CH2)20(CH2)3NH-, H2N(CH2)2NH(CH2)2NH- , H2N(CH2)3NH(CH2)i4 NH-„ H2N(CH2)3 (CH3)(CH2)3NH-, H2N(CH2)2NH(CH2)2.3(CH2)2NH-, H2N(CH2)3NH(CH2)2-4(CH2)2NH(CH2)2NH -, H2N(CH2)3N(CH2 CH2)NH(CH2)3NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)3NH(CH2)CH=CH(CH2)NH(CH2)3NH -.
More specifically this further particular embodiment of the invention is concerned with the use of the compounds for targeting HIV gp120 wherein the compound is selected from the list of
- 1 ,3,5-Tris(2,3-dihydroxybenzamidomethyl)-2,4,6-triethylbenzene (9, PCB311);
- 1 ,3,5-Tris(2,3-dihydroxybenzamidomethyl)-2,4,6-triethylbenzene (10, AL180);
- a- or -D-Glucopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (23, AL-18);
- a- or S-D-Glucopyranose pentakis[3,5-bis(phenylmethoxy)benzoate];
- a- or β-D-Glucopyranose pentakis[3,4-bis(phenylmethoxy)benzoate];
- σ or -D-Glucitol 1 ,5-anhydro-pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j6-D-Galactopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (25, AL-29 and AL-30);
- a- or jS-D-Allopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or -D-Mannopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate](29, 30 and 31);
- a- or j8-D-Xylopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or jS-L-Arabinopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j8-D-Ribopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate] (27, AL36);
- a- or j8-D-Luxopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- σ- or β-D-Glucopyranose pentakis[3,5-dihydroxybenzoate];
- a- or /3-D-Glucopyranose pentakis[3,4-dihydroxybenzoate];
- D-Galactopyranose pentakis[3,4-dihydroxybenzoate];
- D-Arabinopyranose tetrakis[3,4-dihydroxybenzoate];
- D-Mannopyranose pentakis[3,4-dihydroxybenzoate];
- σ or jS-D-Glucitol 1 ,5-anhydro-pentakis[3,4,5-trihydroxybenzoate];
- a- or -D-Glucopyranose pentakis[3,4,5-trihydroxybenzoate] (24, AL-19);
- a- or β-D-Galactopyranose pentakis[3,4,5-trihydroxybenzoate] (26, AL-33);
- a- or -D-Allopyranose pentakis[3,4,5-trihydroxybenzoate];
- a- or -D-Mannopyranose pentakis[3,4,5-trihydroxybenzoate](32, AL51a and 33, AL51b);
- a- or β-D-Xylopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or -L-Arabinopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or jS-D-Ribopyranose tetrakis[3,4,5-trihydroxybenzoate] (28, AL50);
- a- or -D-Luxopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or j8-D-Glucopyranose pentakis[3,4,5-trimethoxybenzoate];
- a- or j8-D-Mannopyranose pentakis[3,4,5-trimethoxybenzoate]
-]8-D-Glucopyranose1-[4,5-dihydroxy-2-(phenylmethoxy)benzoate]2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate];
- β -D-Glucopyranose, 1-[3,4-dihydroxy-5-(phenylmethoxy)benzoate] 2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate].
Another aspect of the invention is a pharmaceutical composition comprising a compound of the invention linked to an HIV inhibitor or antibody-recruiting therapeutic and a proces for preparing such a pharmaceutical composition 9 wherein a compound of the invention is intimately mixed with one or more pharmaceutical excipients or pharmaceutically acceptable carriers.
The invention is further concerned with the above mentioned pharmaceutival compositions for the treatment of an HIV infection or the use of said composition for the manufacture of a medicament for the treatment of an HIV infection
The invention also relates to a separation matrix for affinity chromatography of gp120, which matrix comprises the compounds of the invention.
Another aspect of the present invention relates to a method for (a) binding or purifying (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus, or (3) gp120 comprising viruses or for (b) separating (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus or (3) gp120 comprising viruses, from other proteins, cells or viruses which do not comprise gp120, wherein said method comprises the use of a compound of the invention. The invention also relates to a method for quantifying, titrating or detecting gp120 of fragments thereof (to which the compounds bind).
The invention further provides for the use of the compounds and for methods using the compounds for directing anti-viral drugs or other agents to gp120 comprising viruses or to gp120 comprising virus-infected cells.
Yet another aspect relates to the use of the compounds of the invention for (a) binding or purifying (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus, or (3) gp120 comprising viruses or for (b) separating (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus or (3) gp120 comprising viruses, from other proteins, cells or viruses which do not comprise gp120, wherein said method comprises the use of a compound of the invention. The invention also relates to the use of the compounds of the invention for quantifying, titrating or detecting gp120 of fragments thereof (to which the compounds bind).
DETAILED DESCRIPTION OF THE INVENTION
The present invention will be described with respect to particular embodiments but the invention is not limited thereto.
It is to be noticed that the term "comprising", used in the claims, should not be interpreted as being restricted to the means listed thereafter; it does not exclude other elements or steps.
Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment, but may. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner, as
would be apparent to one of ordinary skill in the art from this disclosure, in one or more embodiments. Where an indefinite or definite article is used when referring to a singular noun e.g. "a" or "an", "the", this includes a plural of that noun unless something else is specifically stated.
Similarly it should be appreciated that in the description of exemplary embodiments of the invention, various features of the invention are sometimes grouped together in a single embodiment, figure, or description thereof for the purpose of streamlining the disclosure and aiding in the understanding of one or more of the various inventive aspects.
The term "virus comprising gp120", "gp120 comprising virus" or "gp120 containing virus" as used herein means a virus which comprises a gp120 gene and which exposes at least parts or fragments of the glycoprotein gp120 on its surface. In a particular embodiment, the term gp120 comprising virus refers to HIV, the human immunodeficienccy virus which has the gp120 gene, env, and which exposes part of gp120 on the surface of the HIV envelope.
The term "associating with" or "binding" refers to a condition of proximity between a chemical entity, or portions thereof, and a target protein or peptide fragment thereof. The association may be non-covalent, wherein the juxta-position is energetically favoured by hydrogen bonding or van der Waals or electrostatic interactions, or alternatively it may be covalent. In each of the following definitions, the number of carbon atoms represents the maximum number of carbon atoms generally optimally present in the substituent or linker; it is understood that where otherwise indicated in the present application, the number of carbon atoms represents the optimal maximum number of carbon atoms for that particular substituent or linker. The term "leaving group" or "LG" as used herein means a chemical group which is susceptible to be displaced by a nucleophile or cleaved off or hydrolyzed in basic or acidic conditions. In a particular embodiment, a leaving group is selected from a halogen atom (e.g., CI, Br, I) or a sulfonate (e.g., mesylate, tosylate, triflate).
The term "protecting group" refers to a moiety of a compound that masks or alters the properties of a functional group or the properties of the compound as a whole. The chemical substructure of a protecting group varies widely. One function of a protecting group is to serve as intermediates in the synthesis of the parental drug substance. Chemical protecting groups and strategies for protection/deprotection are well known in the art. See: "Protective Groups in Organic Chemistry", Theodora W. Greene (John Wiley & Sons, Inc., New York, 1991. Protec''
groups are often utilized to mask the reactivity of certain functional groups, to assist in the efficiency of desired chemical reactions, e.g. making and breaking chemical bonds in an ordered and planned fashion. Protection of functional groups of a compound alters other physical properties besides the reactivity of the protected functional group, such as the polarity, lipophilicity (hydrophobicity), and other properties which can be measured by common analytical tools. Chemically protected intermediates may themselves be biologically active or inactive. Protected compounds may also exhibit altered, and in some cases, optimized properties in vitro and in vivo, such as passage through cellular membranes and resistance to enzymatic degradation or sequestration. In this role, protected compounds with intended therapeutic effects may be referred to as prodrugs. Another function of a protecting group is to convert the parental drug into a prodrug, whereby the parental drug is released upon conversion of the prodrug in vivo. Because active prodrugs may be absorbed more effectively than the parental drug, prodrugs may possess greater potency in vivo than the parental drug. Protecting groups are removed either in vitro, in the instance of chemical intermediates, or in vivo, in the case of prodrugs. With chemical intermediates, it is not particularly important that the resulting products after deprotection, e.g. alcohols, be physiologically acceptable, although in general it is more desirable if the products are pharmacologically innocuous.
The term "alkyl" or "Ci-ie alkyl" as used herein means C1-C18 normal, secondary, or tertiary, linear or cyclic, branched or straight hydrocarbon with no site of unsaturation. Examples are methyl, ethyl, 1 -propyl (n-propyl), 2-propyl (iPr), 1 -butyl, 2-methyl-1 -propyl(i-Bu), 2-butyl (s- Bu), 2-dimethyl-2-propyl (t-Bu), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl- 2-butyl, 3-methyl-1 -butyl, 2-methyl-1 -butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl- 2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, cyclopropylethylene, methylcyclopropylene, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, n-heptyl, n-octyl, n-nonyl, n- decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecyl, n-hexadecyl, n-heptadecyl, n- octadecyl, n-nonadecyl, n-icosyl, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. In a particular embodiment, the term alkyl refers to C1-12 hydrocarbons, yet more in particular to C1-6 hydrocarbons as further defined herein above.
The term "acyclic alkyl" as used herein means C1-C18 normal, secondary, or tertiary, linear, branched or straight, hydrocarbon with no site of unsaturation. Examples are methyl, ethyl, 1 -propyl, 2-propyl (iPr), 1 -butyl, 2-methyl-1-propyl(i-Bu), 2-butyl (s-Bu), 2-methyl-2-propyl (t-Bu), 1-pentyl (n-pentyl), 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1 -butyl, 2- methyl-1 -butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pe
3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecyl, n-hexadecyl, n- heptadecyl, n-octadecyl, n-nonadecyl and n-icosyl.
The term "cycloalkyl" or "C3-18 cycloalkyl" as used herein and unless otherwise stated means a saturated hydrocarbon monovalent radical having from 3 to 18 carbon atoms consisting of or comprising a C3-10 monocyclic or CMS polycyclic saturated hydrocarbon, such as for instance cyclopropyl, cyclobutyl, cyclopentyl, cyclopropylethylene, methylcyclopropylene, cyclohexyl, cycloheptyl, cyclooctyl, cyclooctylmethylene, norbornyl, fenchyl,
trimethyltricycloheptyl, decalinyl, adamantyl and the like.
The term "alkenyl" or "CMsalkenyl" as used herein is C2-C18 normal, secondary or tertiary, linear or cyclic, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond. Examples include, but are not limited to: ethylene or vinyl (-CH=CH2), allyl (-CH2CH=CH2), cyclopentenyl (-C5H7), cyclohexenyl (-C6H9), cyclopentenylpropylene, methylcyclohexenylene and 5-hexenyl (- CH2CH2CH2CH2CH=CH2). The double bond may be in the cis or trans configuration. In a particular embodiment, the term alkenyl refers to C1.12 hydrocarbons, yet more in particular to C1-6 hydrocarbons as further defined herein above.
The term "acyclic alkenyl" as used herein refers to C2-Ci8 normal, secondary or tertiary, linear, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond. Examples include, but are not limited to: ethylene or vinyl (-CH=CH2), allyl (-CH2CH=CH2) and 5-hexenyl (-CH2CH2CH2CH2CH=CH2). The double bond may be in the cis or trans configuration.
The term "cycloalkenyl" as used herein refers to a non-aromatic hydrocarbon radical having from 3 to 18 carbon atoms with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond and consisting of or comprising a C3-10 monocyclic or C7-18 polycyclic hydrocarbon. Examples include, but are not limited to:
cyclopentenyl (-C5H7), cyclopentenylpropylene, methylcyclohexenylene and cyclohexenyl (-C6H9). The double bond may be in the cis or trans configuration.
The term "alkynyl" or "CMsalkynyl" as used herein refers to C2-C18 normal, secondary, tertiary, linear or cyclic, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp triple bond. Examples include, but are not limited to: ethynyl (-C≡CH), 3-ethyl-cyclohept-1 -ynylene, 4-cyclohept-1-yn-methylene and 1- propynyl (propargyl, -CH2C≡CH). In a particular embodiment, the term alkenyl refers to C1-12 hydrocarbons, yet more in particular to C1-6 hydrocarbons as further defined herein above.
The term "acyclic alkynyl" as used herein refers to C2-C18 normal, secondary, tertiary, linear, branched or straight hydrocarbon with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp triple bond. Examples include, but are not limited to: ethynyl (-C≡CH) and 1-propynyl (propargyl, -CH2OCH).
The term "cycloalkynyl" as used herein refers to a non-aromatic hydrocarbon radical having from 3 to 18 carbon atoms with at least one site (usually 1 to 3, preferably 1 ) of unsaturation, namely a carbon-carbon, sp triple bond and consisting of or comprising a C3-10 monocyclic or C7-18 polycyclic hydrocarbon. Examples include, but are not limited to: cyclohept-1- yne, 3-ethyl-cyclohept-1-ynylene, 4-cyclohept-1-yn-methylene and ethylene-cyclohept-1-yne.
The term "alkylene" as used herein each refer to a saturated, branched or straight chain hydrocarbon radical of 1-18 carbon atoms (more in particular Ci-i2 or Ci-s carbon atoms), and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkane. Typical alkylene radicals include, but are not limited to: methylene (-CH2-), 1 ,2-ethyl (-CH2CH2-), 1 ,3-propyl (-CH2CH2CH2-), 1 ,4-butyl (- CH2CH2CH2CH2-), and the like.
The term "alkenylene" as used herein each refer to a branched or straight chain hydrocarbon radical of 2-18 carbon atoms (more in particular C2-i2 or C2-6 carbon atoms) with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp2 double bond, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkene.
The term "alkynylene" as used herein each refer to a branched or straight chain hydrocarbon radical of 2-18 carbon atoms (more in particular C2-i2 or C2-6 carbon atoms) with at least one site (usually 1 to 3, preferably 1) of unsaturation, namely a carbon-carbon, sp triple bond, and having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkyne.
The term "heteroalkyl" as used herein refers to an acyclic alkyl wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom. The term heteroalkyl thus comprises -O-alkyl, -NH-alkyl, -N(alkyl)2, and -S-alkyl.
The term "heteroalkenyl" as used herein refers to an acyclic alkenyl wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom. The term heteroalkenyl thus comprises -O-alkenyl, -NH-alkenyl, -N(alkenyl)2, -N(alkyl)(alkenyl), and -S-alkenyl.
The term "heteroalkynyl" as used herein refers to an acyclic alkynyl wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom. The term heteroalkynyl tl
comprises -0-alkynyl, -NH-alkynyl, -N(alkynyl)2, -N(alkyl)(alkynyl), -N(alkenyl)(alkynyl), and -S- alkynyl.
The term "heteroalkylene" as used herein refers to an alkylene wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
The term "heteroalkenylene" as used herein refers to an alkenylene wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom.
The term "heteroalkynylene" as used herein refers to an alkynylene wherein one or more carbon atoms are replaced by an oxygen, nitrogen or sulphur atom. As used herein and unless otherwise stated, the term halogen means any atom selected from the group consisting of fluorine (F), chlorine (CI), bromine (Br) and iodine (I).
Any substituent designation that is found in more than one site in a compound of this invention shall be independently selected.
Substituents optionally are designated with or without bonds. Regardless of bond indications, if a substituent is polyvalent (based on its position in the structure referred to), then any and all possible orientations of the substituent are intended.
Detailed description The present invention relates to a series of novel compounds which have been shown to have strong binding or association capacity to the glycoprotein gp120. The invention therefore relates to the new compounds, methods for their preparation, the use of the compounds for binding, purifying, titrating (quantifying), removing or separating gp120 and fragments thereof, cells infected with gp120 comprising viruses or viruses comprising gp120 and for targeting antiviral drugs or other substances or agents to gp120 (such as to gp120 comprising viruses).
The present invention provides novel compounds which have a central benzene scaffold or a central carbohydrate scaffold, with a general structure according to the formulas (A), (B), (C), (D) or (E),
(C) (D) (E)
wherein,
- each n is independently selected from 1 and 2;
- L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene,
heteroalkenylene, and heteroalkynylene can be unsubstituted or substituted with =0, =S; OH; - SH; and NH2;
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
— N—
heteroalkynylene; and (1 ,4-piperazinylene);
- each of R1a, R1b and R1c is independently selected from methyl; and ethyl;
- each of R, R3, R4 and R5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3; or
- one of X-R3, X-R4 or X-R5 is a linking moiety to which another substance or agent can be attached.
In a particular embodiment, the linking moiety is selected from C 1 -1 oo etero al ky I ; Ci- looheteroalkenyl; and Ci-iooheteroalkynyl, wherein said Ci-iooheteroalkyl, Ci-iooheteroalkenyl and Ci-iooheteroalkynyl can be unsubstituted or substituted with one or more =0, =S; OH; -SH; and NH2.
The compounds of the invention can have a central benzene scaffold or a central carbohydrate scaffold and a variation of compounds with these central scaffolds are envisaged in this invention. Compounds of formula AA containing 2,4,6-triethyl(methyl)benzene as central scaffold are envisaged with this invention. For their preparation, first, di-and trihydroxy benzoyl I and caffeoyl II moieties are introduced and in addition, some non aromatic polyhydroxy residues structurally related to I such as shikimyl III or quinyl IV are introduced. These residues are connected to the central scaffold by ester or amide bonds.
verview 1 : compounds of formula AA
Compounds of formula AA which are directly connected to the amino nitrogen of aniline (V) or benzylamine (VI) moieties substituted with two or three OH are envisaged herein. Overview 2
Ri = Me, Et
F¾ = R3 = R4
R2 = R3 F¾
R2 ^ R3 ^ R4
Moreover, compounds of general formula BB in which two 2,4,6-triethyl(methyl)benzene scaffolds are linked through spacers of different length and nature, are also envisaged (Overview 3).
Overview 3: Compounds with formula BB
2
n = 2, X = NH
Compounds of general formula CC and DC with monosaccharides as central scaffolds and disaccharides (general formula EE) are also envisaged (Overview 4). The different stereochemistry and ring sizes, of these "scaffolds" offer a unique opportunity to modify the number and distribution of the groups attached to the central scaffold.
Overview 4: Compounds with carbohydrates as central scaffold
Compounds can furthermore be linked (i.e. directly or through a spacer molecule) to a therapeutic drug or to a dye, a fluorescent molecule, a diagnostic enzyme, or a radiolabeled entity to enable identification of gp120 containing viruses or virus-infected (gp120 expressing) cells or to target therapeutically gp120 containing viruses or virus-infected (gp120 expressing) cells. When the compounds are bound to a column or to beads, they can allow to selectively bind, identify and/or remove gp120, gp120-containing viruses or gp120-expressing cells, from media, and biological fluids (i.e. blood, sera, plasma), and they can allow isolation and purification (or enriching) of gp120 and/or gp120-containing viruses and gp120-expressing cells.
Linking or coupling the compounds of the invention to a therapeutic drug or other agents or substances can be achieved with methods known to the person skilled in the art and examples are described herein.
The compounds of the invention can thus be used to bind, purify, titrate, quantify, remove or separate gp120 or fragments thereof, cells infected with gp120 comprising viruses or viruses comprising gp120 and can be used for targeting antiviral drugs or other substances or agents to gp120 (such as to gp120 comprising viruses).
Hence another aspect of the invention relates to a pharmaceutical composition comprising a compound of the invention as a carrier for targeting antiviral drugs or other substances or agents to gp120 (such as to gp120 comprising viruses or cells). The compositions of the invention may be administered to the mammal (including a human) to be treated by any means well known in the art, i.e. orally, intranasally, subcutaneously, intramuscularly, intradermal^, intravenously, intra-arterially, parenterally or by catheterization.
The invention also relates to a process of preparing a pharmaceutical composition of the invention wherein the compounds of the invention, carrying the one or more therapeutical antiviral drugs or other substances or agents and the optionally one or more pharmaceutical excipients or pharmaceutically acceptable carriers are intimately mixed.
The invention further relates to the use of a composition comprising (a) one or more derivatives of formula (A), (B), (C), (D) or (E) as carrier for targeting one or more viral inhibitors to provide a synergistic effect against a viral infection, preferably a HIV/AIDS viral infection. Within the framework of this embodiment, the viral inhibitors may be
• (Co)receptor inhibitors [i.e. maraviroc (CCR5), bicyclams such as AMD3100 (CXCR4), CD4- binding agents]
• Carbohydrate-binding agents (CBA) (i.e. pradimicins, lectins, 2G12)
• Specific gp120 or gp41 binding compounds i.e. BMS806, T20)
· Anionic compounds (i.e. PRO-2000, PVAS, PAVAS)
Within the framework of this embodiment, the viral inhibitors or other substances or agents may be antibody-recruiting therapeutics such as but not limited to:
-dinitrophenyl(DNP),
- -Gal conjugates.
The bifunctional molecule that is obtained by coupling the compounds of the inventions to the above mentioned antibody-recruiting therapeutics can recruit antibodies such as but not limited to:
-anti-DNP antibodies
-human anti-Gal antibodies
to gp120-expressing cells and initiate an immune respons (such as but not limited to a functional complement-dependent cytotoxic response) against gp120-expressing cells. Furthermore these bifunctional molecules can bind competitively to gp-120-expressing cells and inhibit f.e. gp120- CD4 interactions, thus decreasing viral infectivity (Parker et al. 2009, J. Am. Chem. Soc. 131 , 16392-16394; Naicker et al. 2004, 2, 660-664; Perdome et al. 2008, Proc. Natl. Acad. Sci. U.S.A.105, 6). The invention also relates to the compositions of the invention being used for inhibition of the replication of HIV strains.
More generally, the invention relates to the compositions of the invention being useful as agents having biological activity (preferably antiviral activity) or as diagnostic agents. Any of the uses mentioned with respect to the present invention may be restricted to a non-medical use, a non- therapeutic use, a non-diagnostic use, or exclusively an in vitro use, or a use related to cells remote from an animal.
The invention thus relates to a method for the treatment of viral infections in humans and other mammals wherein a therapeutically effective amount of the composition is administered to a human or a mammal in need of said treatment. The therapeutically effective amount, especially for the treatment of viral infections in humans and other mammals, is a HIV protein inhibiting amount. More particularly, it is a HIV replication inhibiting amount. As an example, the compounds of the invention can be linked to or associated with (absorbed on or into) a polymer or a matrix used in column chromatography, such as in affinity chromatography in order to create a new matrix for column chromatography for binding, separating or purifying gp120 or fragments thereof.
Thus, in a specific embodiment, the compounds of the invention is to be used in a form immobilised to a solid support, e.g. as a ligand in affinity chromatography for gp120, wherein the compounds or a linker attached thereto comprises terminating functionalities useful for such immobilisation.
Another embodiment relates to a separation matrix for use in affinity chromatography, wherein the ligands of the separation mix comprise at least one compound as defined above. In a specific embodiment, the ligands have been coupled to a support via linkers. The present matrix can e.g. be in the form of separate particles, preferably porous and essentially spherical particles; a monolith; or a membrane. Also encompassed by the invention is a system suitable for affinity chromatography, which is comprised of a separation matrix as defined above packed in a column. The column may be of a size suitable for analytical scale or for large scale chromatography.
Suitable support materials are well known. In one embodiment, the support is a natural polymer, such as agarose, alginate, carrageenan, gelatine etc. Such natural polymers are known to form physically cross-linked networks spontaneously on cooling or on addition of divalent metal ions, and chemical cross-linkers can be added if desired. This kind of supports is easily prepared according to standard methods, such as inverse suspension gelation (S Hjerten, Biochim Biophys Acta 79(2), 393-398 (1964)). In another embodiment, the support is comprised of cross-linked synthetic polymers, such as styrene or styrene derivatives, divinylbenzene, acrylamides, acrylate esters, methacrylate esters, vinyl esters, vinyl amides etc. Such polymers are also easily produced according to standard methods, see e.g. "Styrene based polymer supports developed by suspension polymerization" (R Arshady: Chimica e L'lndustria 70(9), 70-75 (1988)). Thus, in summary, the support material can in principle be any material that allows the covalent coupling of the gp120 binding compounds of the invention, such as the above-discussed polymers, inorganic materials, such as silica, ceramics etc.
Many well-known methods are available for immobilising ligands to a support through suitable functional groups. As the skilled person in this field will realise, the exact choice of coupling method will depend on the structure of the ligand to be immobilised. In one embodiment, the support has hydrophilic surfaces, and if porous, the surfaces of the pores are also hydrophilic. This is advantageous in order to avoid or at least reduce any non-specific protein interactions. It is also advantageous if the surfaces have a high density of groups available for coupling of ligands. Such coupling groups are commonly hydroxyl groups, but may also be ally, groups i.e. double bonds available for grafting, amines, thioles, epoxides and the like. If the support material has undesirable surface properties, it is possible to coat it with a hydrophilic polyhydroxy- functional material before coupling the ligand. The techniques and considerations for coupli
affinity ligands to a suitable support to prepare a separation matrix are well known in this field, see e.g. WO 98/33572 for a detailed review of coupling chemistry as well as suitable linking molecules.
As described above, the compounds can be used in a method of isolating or separating gp120 or fragments thereof, from other components, for example in a liquid, wherein a compound or a separation matrix as defined above is used. In the most preferred embodiment, the present method is affinity chromatography, which is a widely used and well-known separation technique. In brief, in a first step, a solution comprising gp120 or binding fragments thereof is passed over a separation matrix under conditions allowing adsorption of gp120 or fragments thereof to the compounds of the present invention present on said matrix. Such conditions are controlled e.g. by pH and/or salt concentration i.e. ionic strength in the solution. Care should be taken not to exceed the capacity of the matrix, i.e. the flow should be sufficiently slow to allow a satisfactory adsorption. In this step, other components of the solution will pass through in principle unimpeded. Optionally, the matrix is then washed, e.g. with an aqueous solution, in order to remove retained and/or loosely bound substances. In a next step, a second solution denoted an eluent is passed over the matrix under conditions that provide desorption i.e. release of gp120 or fragments thereof. Such conditions are commonly provided by a change of the pH, the salt concentration i.e. ionic strength, hydrophobicity etc. Various elution schemes are known, such as gradient elution and step-wise elution. Elution can also be provided by a second solution comprising a competitive substance, which will replace the gp120 or fragments thereof on the matrix.
In an alternative embodiment, the compounds of the invention is used in an assay for detection of gp120, fragments thereof or gp120 comprising cells. In this case, the compound is preferably labelled with a suitable detectable label as conventionally used, such as a fluorescent label, a luminescent label, a chemiluminiscent label, an enzyme label, a radioactive label, an absorbance label etc.
Alternatively, the compounds of the invention can be linked to anti-HIV entry inhibitors in order to target to the anti-HIV drugs to the site of interaction and to retain the anti-HIV drugs for a longer period at their site of interaction.
The compounds of the invention according to the formulae of the application can be prepared while using a series of chemical reactions known to those skilled in the art, altogether making up the process for preparing said compounds and exemplified further. The proce
described further are only meant as examples and by no means are meant to limit the scope of the present invention.
Compounds with a Benzene scaffold.
The synthesis of the benzyl protected derivatives (5 or PCB245) and (7 or AL-168) was accomplished by coupling of the trisamine (1) (Walsdorff, C. et al. J. Chem.Res (M) 1996, 1601- 1609; Metzger, A., et al. Angew. Chem. 1997, 109, 911-914; Metzger, A. et al. Angew. Chem Ed. Engl. 1997, 36, 862-864; Wallace, K. J. et al. Synthesis 2005, 12, 2080-2083) with the respective benzyl protected trihydroxybenzoic acids (2) (Belin, F. et al. Helv. Chim. Acta 2003, 86, 247-265) and (3) (JP 08143525 A) in the presence of BOP and triethylamine (Scheme 1). Reaction of (1) with the commercially available 2,3-dimethoxybenzoic acid (4) under the same coupling conditions afforded (9) (PCB311) (Stack, T.D.P. et al. J. Am. Chem. Soc. 1993, 115, 6466-6467) in which the phenol groups were protected as methyl ethers. The preparation of analogs of 9 and 10 wherein the ethylgroups on the benzylscaffold are replaced with methylgroups are also described (Hou Z; Inorganic Chemica Acta (1997), 263 (1-2), 341-355.
cheme 1
To prepare compounds in which the polyphenol moiety was linked directly to the benzene scaffold (Scheme 2), the 1 ,3,5-tris(bromomethyl)-2,4,6-trietylbenzene intermediate 11 (Walsdorff, C. et al. J. Chem.Res (M) 1996, 1601-1609; Metzger, A., et al. Angew. Chem. 1997, 109, 911- 914; Metzger, A. et al. Angew. Chem Ed. Engl. 1997, 36, 862-864; Wallace, K. J. et al. Synthesis 2005, 12, 2080-2083) was used as starting material. Treatment of 11 with the commercially available amines: 3,4-dimethoxyaniline (12), 3,4,5-trimethoxyaniline (13) and 3,4,5- trimethoxybenzylamine (14) in the presence of triethylamine or potassium carbonate afforded the methoxy protected derivatives: 15, 17 and 18 (AL125, AL126 and AL177) respectively. Deprotection of 15 (AL125) and 18 (AL177) with boron tribromide gave the corresponding OH deprotected derivatives 16 (AL172) (94%) and 19 (AL178) (99%) in excellent yields.
Another example is provided in Scheme 3. Reaction of the benzoic acid 3 with ethylenediamine afforded the A/-(2-aminoethyl)-2,3,4-tris(benzyloxy)benzamide 20. Reaction of 20 with the tris(bromomethyl) derivative 11 in the presence of trietylamine gave 21 (ECMVI27) which was deprotected by hydrogenation to afford 22 (AL184). Scheme 3
ECM-VI-27; R= Bn
H2 (
Pd/CV 22 AL-184; R= H
Compounds with a carbohydrate scaffold.
The a and β anomer of penta-O-galloyl-D-glucopyranose (compound 24) exist in nature (Li, Y, et al. Biochem Biophys. Res. Commun. 2005, 336, 430-437). The synthesis of these compounds has been described in Khanbabaee, K. et al. Tetrahedron 1997, 53, 10725-10732 and is schown in scheme 4 and partially in scheme 5. The synthesis of the benzyl protected glucose, galactose, ribose and mannose derivatives (23, 25, 27, 30 and 31) (Scheme 4) have been described in Ren, Y. et al. J.Med. Chem. 2006, 49, 2829-2837. The corresponding deprotected compounds (26, 28, 32 and 33) (Scheme 4) have been also described in Ren, Y. et al. J.Med. Chem. 2006, 49, 2829-2837. Also the dihydroxybenzoate, trimethoxybenzoate and bis(phenylmethoxy)benzoate analogues of some of these compounds have been described (Ren, Y. et al. J.Med. Chem. 2006, 49, 2829-2837, Barres T; et al. Carbohydrate Research (2004) 339(7), 1373-1376 and US 20100137194 published on 3 June, 2010). Some of these compounds have been described to act as insulin mimetics stimulating glucose transport in adipocytes and reducing blood glucose and insulin levels in diabetic and obese animals. Other compounds have been described as plasminogen activator 1 inhibitors.
Scheme 4
D-glucose 23 (AM 8) 24 (AL-19)
D -galactose 25 (AL29 + AL30) 26 (AL33)
D-ribose 27(AL36) 28 (AL50)
20100137194 published on 3 June, 2010).
Commercially available D-mannose was allowed to react with 2,3,4 tribenzylbenzoic acid 311 in the presence of DMAP and DCC to afford the corresponding benzyl protected derivative 34 as a mixture of a and β anomers (Scheme 5). Separation of the anomers by column chromatography on silica gel using a mixture of toluene/ethyl acetate (96:4) as eluent gave the a 35 and β 36 anomers in 33 % and 20 % yields respectively. Debenzylation of the individual anomers using hydrogenation conditions gave the corresponding deprotected derivatives 37 (78%) and 38 (80%) (Scheme 5).
39
Linking of gp120 binding compounds to other agents or substances
As described herein, the gp120 binding compounds can be linked (whether or not via a linker) to a therapeutic drug, on a column polymer, to a dye, a fluorescent molecule, a diagnostic enzyme, or a radiolabeled entity to enable identification of gp120 containing viruses or virus-infected. The synthesis of a gp120 binding compound of the invention with a linker moiety can be performed as shown in scheme 7. Subsequently, the therapeutic drug, polymer, dye, fluorescent molecule, diagnostic enzyme, or radiolabeled entity, amongh others can be coupled to the linker through reactions known in the art.
Examples
EXAMPLE 1 : GENERAL SYNTHETIC METHODS AND MATERIALS FOR THE PREPARATION OF THE COMPOUNDS WITH A CENTRAL BENZENE SCAFFOLD
General Methods.
The starting materials for the synthesis were used as purchased from Sigma-Aldrich. 1H NMR and 13C NMR spectra were recorded on Varian lnova-300 (300 and 75 MHz, respectively), Varian lnova-400 (400 and 100 MHz, respectively), Bruker-300 (300 and 75 MHz, respectively) and Bruker-500 (500 MHz) instruments with the solvent signals as internal standard. Ion mass spectra were performed on a Hewelett Packard 1100 MSD, on a Micromass ZQ and on an Agilent Technologies 6520 Accurate-Mass QTOF LC/MS. For HPLC analysis was used an Alliance 2695 (Waters) equipped with a PDA (Photo Diode Array) detector Waters 2996. Acetonitrile was used as mobile phase A with 0.08% of formic acid, and water was used as mobile phase B with n λ η'
of formic acid at a flow rate of 1 ml/min. Two different methods were used, one on a XBridge Cie (2.1 x 100 mm, 3.5 μΐτι) column and A:B, 5-80% of A that will be noted as f/?pg and the other on a SunFire Cie (4.6 x 50 mm, 3.5 μΐτι) column with a A:B, 0-100% of A, that will be noted as t^s). Melting points were measured on a Reichert-Jung Kofler. Optical rotations were measured on a Perkin-Elmer 241 MC polarimeter with a cell of path length 1 dm.
Synthetic procedures for some of the compounds can be found in:
Walsdorff, C. etal, J. Chem.Res (M) 1996, 1601-1609;
Metzger, A. et al, Angew. Chem. 1997, 109, 911-914;
Metzger, A. et al, Angew. Chem Ed. Engl. 1997, 36, 862-864;
Wallace, K. J. et al, Synthesis 2005, 12, 2080-2083;
Belin, F. et al, Helv. Chim. Acta 2003, 86, 247-265; and
JP 08143525 A. General procedure A for the reaction of trisamine 1 with benzoic acids (in accordance with Scheme 1). Benzotriazol-1-yloxytris(dimethylamino)phosphoniumhexafluorophosphate (BOP) (3.3 eq) was added to a solution of the corresponding benzoic acid (3.3 eq) in dry dichloromethane (5 mL). After 5 min, trisamine 1 (50 mg, 1 eq) and triethylamine (3.3 eq) was added. The mixture was stirred at room temperature overnight. The solvent was evaporated and the residue was treated with ethyl acetate (10 mL) and washed successively with saturated solutions of citric acid (3x 30 mL), NaHC03 (3 x 30 mL) and brine (1 x 30 mL). The organic phase was dried (Na2S04), filtered and evaporated to dryness. The residue was purified by CCTLC on the chromatotron using (dichlorometane/methanol, 9:1) as eluent. General procedure B for the reaction of trisbromomethyl derivative 11 with amines (in accordance with Scheme 2). To a solution of 11 (1 eq) and the corresponding amine (4 eq) in dichloromethane (5 mL) was added triethylamine (4 eq), and the mixture was stirred at room temperature overnight. Dichloromethane (25 mL) was added and the reaction mixture was washed with a saturated solution of ammonium chloride (3 x 25 mL). The organic layer was dried (Na2S0 ), filtered, and evaporated to dryness. The residue was purified by CCTLC.
General procedure C for benzyl group deprotection. A solution of the corresponding benzyl protected derivative (1 mmol) in THF/methanol (1 :1) (100 mL) containing 30 wt% of Pd/C (10%) was hydrogenated at 2.85 atm (42 psi) at 30 °C overnight. The Pd/C was filtered thi
Whatman® filter paper 42 and the solvent was removed under reduced pressure to give the corresponding deprotected derivatives as unique products.
General procedure D for methyl group deprotection. The starting O-methylated compound (1 mmol) was dissolved in dry dichloromethane (10 ml_). The solution was cooled to 0° C, and BBr3 (1.5 eq for each OMe) was added under argon atmosphere. The mixture was allowed to slowly reach room temperature. After completion of the reaction (16-18 h) a white precipitate was formed. The solid was filtered and tritutared with cool dichloromethane and ethyl ether to give the pure deprotected compound.
EXAMPLE 2: Synthesis of 1,3,5-Tris(3,4,5-tribenzyloxybenzamidomethyl)-2,4,6- triethylbenzene (5, PCB245). Trisamine derivative 1 (50 mg, 0.2 mmol) and 2 (317 mg, 0.72 mmol) was reacted according to the general procedure A to give 212 mg (70%) of 5, PCB245 as a white solid: mp 178-180 °C. HPLC: tRQq = 20.44. 1H NMR (CDCI3, 300 MHz) δ: 1.18 (t, J = 6.9 Hz, 9H, CH3), 2.73 (q, J 6.7 Hz, 6H, CH3-CH2), 4.62 (s, 6H, CH2-NH), 4.99 (s, 18H, CH2-Ph), 5.69 (s, 3H, NH), 6.93 (s, 6H, H-Ar), 7.14-7.30 (m, 45H, H-Ar). C NMR (CDCI3, 50 MHz) δ: 17.15 (CH3), 23.76 (CH2), 39.17 (CH2), 72.09 (CH2), 75.63 (CH2), 107.76 (CH), 127.96-128.99 (CH), 132.78 (C), 136.95 (C), 137.67 (C), 142.11 (C), 145.21 (C), 153.25 (C), 167.20 (CO). MS (ES+) m/z: 1517 (M+H)+, 1539 (M+Na)+.
EXAMPLE 3: Synthesis of 1,3,5-Tris(3,4,5-trihydroxybenzamidomethyl)-2,4,6- triethylbenzene (6, AL 113). Following the general procedure C, the benzyl derivative 5, PCB245 (100 mg, 0.066 mmol) gave 46 mg (99 %) of 6, AL 113 as a white solid: mp 178-180 °C. HPLC: tR(xj = 3.04 min. 1H NMR (CD3OD, 300 MHz) δ: 1.21 (t, J = 7.3 Hz, 9H, CH3), 2.90 (q, J = 7.1 Hz, 6H, CH3-CH2), 4.61 (s, 6H, CH2-NH), 6.81 (s, 6H, H-Ar). EM (ES+): m/z 706 (M+H)+, 728 (M+Na)+.
EXAMPLE 4: Synthesis of 1,3,5-Tris(2,3,4-tribenzyloxybenzamidomethyl)-2,4,6- triethylbenzene (7, AL 168). Trisamine derivative 1 (50 mg, 0.2 mmol) and 3 (290 mg, 0.66 mmol) was reacted according to the general procedure A to give 190 mg (63 %) of 7, AL 168 as a yellow oil. HPLC: tR(S) = 5.74 min. Ή NMR (CDCI3, 300 MHz) δ: 1.03 (t, J = 7.3 Hz, 9H, CH3- CH2), 2.47 (q, J = 7.3 Hz, 6H, CH3-CH2), 4.53 (s, 6H, CH2-NH), 4.82 (s, 6H, CH2-Ph), 4.83 (s, 6H, CH2-Ph), 5.12 (s, 6H, CH∑-Ph), 6.8-7.4 (m, 48H, H-Ar), 7.74 (t, J = 4.4 Hz, 3H, NH), 7.9 (d, J = 8.9 Hz, 3H, H-Ar). «C NMR (CDCI3, 75 MHz) δ 16.27 (CH3), 22.83 (CH2), 37.88 (CH2), 70.87 (CH2),
75.52 (CH2), 76.48 (CH2), 109.25 (C), 119.68 (C), 126.77 (CH), 127.54-128.06 (CH), 132.34 (C), 135.73 (C), 136.14 (C), 136.88 (C), 141.05 (C), 143.90 (C), 151.42 (C), 155.72 (C), 164.42 (C=0). EM (ES+) m/z: 1517 (M+H)+, 1539 (M+Na)+. EXAMPLE 5: Synthesis of 1,3,5-Tris(2,3,4-trihydroxybenzamidomethyl)-2,4,6- triethyl benzene (8, AL 170). Following the general procedure C, the benzyl derivative 7, AL-168 (190 mg, 0.12 mmol) gave 67 mg (76 %) of 8, AL 170 as a white solid: mp 154-156 °C. HPLC: tR(s) = 3.62 min. 1H NMR (CD3OD, 300 MHz) δ: 1.21 (t, J = 7.3 Hz, 9H, CH3-CH2), 2.88 (q, J = 7.0 Hz, 6H, CH3-CH2), 4.67 (s, 6H, CH2-NH), 6.33 (d, J = 8.8 Hz, 3H, H-Ar) , 7.19 (d, J = 8.8 Hz, 3H, H-Ar). "a NMR (CD3OD, 75 MHz) δ: 16.64 (CH3), 24.04 (CH2), 39.15 (CH2), 108.10 (C), 109.48 (CH), 120.37 (CH), 132.87 (C), 133.75 (C), 145.84 (C), 150.45 (C), 150.67 (C), 170.74 (C=0). EM (ES+) m/z: 706 (M+H)+, 728 (M+Na)+. Anal. Calcd. for C36H39N3O12: C, 61.27; H, 5.57; N, 5.95. Found: C, 60.98; H, 5.63; N, 6.12. EXAMPLE 6: Synthesis of 1,3,5-Tris(3,4-dimethoxyphenylaminomethyl)-2,4,6- triethylbenzene (15, AL-125). To a solution of 11 (100 mg, 0.22 mmol) and the commercially available 3,4-dimethoxyaniline 12 (139.4 mg, 0.91 mmoles) in acetonitrile (6 mL) was added dry potassium carbonate (125.8 mg, 0.91 mmol) and the mixture was refluxed overnight. After evaporation of the solvent, the residue was dissolved in ethyl acetate (15 mL) and washed with water (3 x15 mL). The organic layer was dried (Na2S04), filtered, and evaporated to dryness. The residue was purified by CCTLC (hexane/ethyl acetate, 3:7) to give an orange foam that was treated with ethylacetate/hexane to give 0.093 g (66%) of 15, AL125 as a white solid: mp 77-79 °C. HPLC: tR(xj = 11.57 min. Ή NMR (CDCI3, 300 MHz) δ: 1.26 (t, J = 7.2 Hz, 9H, CH3), 2.79 (q, J = 7.2 Hz, 6H, CH3-CH2), 3.83 (s, 9H, CH3O), 3.86 (s, 9H, CH3O), 4.20 (s, 6H, CH2NH), 6.26 (m, 6H, H-Ar), 6.82 (d, J = 8.3 Hz, 3H, H-Ar) 3C NMR [CDC , 75 MHz] δ: 17.45 (CH3), 23.24 (CH2), 43.35 (CH2), 56.22 (CH3), 57.21 (CH3), 99.14 (CH), 103.35 (CH), 113.76 (CH), 133.77 (C), 142.23 (C), 143.62 (C), 144.02 (C), 150.56 (C). MS (ES+) m/z: 658 (M+H)+.
EXAMPLE 7: Synthesis of 1,3,5-Tris(3,4-dihydroxyphenylaminomethyl)-2,4,6- triethylbenzene (16, AL-172). Following the general procedure D, the methoxy derivative 15, AL125 (50 mg, 0.07 mmol) gave 40 mg (99 %) of 16, AL172 as a white solid: mp >150 °C (dec). HPLC: tR(x) = 10.30 min. Ή NMR (CD3OD, 300 MHz) δ: 1.17 (t, J = 7.3 Hz, 9H, CH3), 2.82 (q, J = 7.3 Hz, 6H, CH3-CH2), 4.66 (s, 6H, CH2-NH), 6.93-7.03 (m, 6H, H-Ar), 6.82 (m, 3H, H-Ar). 3C NMR (CD3OD, 100 MHz) δ 15.95 (CH3), 26.18 (CH2), 52.0 (CH2), 110.82 (CH), 111.10 "
116.96 (CH), 128.44 (C), 128.70 (C) 147.80 (C), 148.35 (C), 149.90 (C). MS (ES+) m/z: 574 (M+H)+. Anal. Calcd. for C33H39N3O6: C, 69.09; H, 6.85; N, 7.32. Found: C, 68.89; H, 6.63; N, 7.43. EXAMPLE 8: Synthesis of 1,3,5-Tris(3,4,5-trimethoxyphenylaminomethyl)-2,4,6- triethylbenzene (17, AL 126). A solution of 11 (150 mg, 0.34 mmol) and the commercially available 3,4,5-trimethoxyaniline 13 (249.2 mg, 1.3 mmol) in acetonitrile (9 mi) was added dry potassium carbonate (188 mg, 1.3 mmol) and the mixture was refluxed overnight. After evaporation of the solvent, the residue was dissolved in ethyl acetate (15 imL) and washed with water (3 x15 mi). The organic layer was dried (Na2S04), filtered, and evaporated to dryness. The residue was purified by CCTLC (ethyl acetate/hexane, 3:7) to give 0.091 g (41%) of 17, AL126 as a white solid: mp 94-96 °C. HPLC: tm = 11.49 min. Ή NMR (CDCI3, 300 MHz) δ : 1.27 (t, J = 7.2 Hz, 9H, CH3-CH2), 2.80 (c, J = 7.2 Hz, 6H, CH3-CH2), 3.79 (s, 9H, m-CH3-0), 3.86 (s, 18H, p- CH3-O), 4.21 (s, 6H, CH2-NH), 5.92 (s, 6H, H-Ar).«C NMR (CDCI3, 75 MHz) δ: 17.45 (CH3), 23.27 (CH2), 43.00 (CH2), 56.48 (CH3), 61.54 (CH3), 90.53 (CH), 130.83 (C), 133.51 (C), 144.16 (C), 145.25 (C), 154.52 (C). EM (ES+) m/z: 748 (M+H)+.
EXAMPLE 9: Synthesis of 1,3,5-Tris(3,4,5-trimethoxybenzylaminomethyl)-2,4,6- triethylbenzene (18, AL 177). A solution of 11 (167 mg, 0.37 mmol) and the commercially available 3,4,5-trimethoxybenzylamine 14 (262 mg, 1.3 mmol) was treated according to the general procedure B. The residue was purified by CCTLC (dichloromethane/methanol/NH4OH, 9:1 :0.2) to give 0.19 g (65%) of 18, AL 177 as a white solid: mp 100-102 °C. HPLC: fow = 2.78. 1H NMR (CDCI3, 300 MHz) δ: 1.11 (t, J = 7.4 Hz, 9H, CH3-CH2), 2.71 (q, , J = 7.4 Hz, 6H CH3- CH2), 3.83 (s, 6H, CH2-NH), 3.87 (bs, 6H, CH3-O and CH2-NH), 6.64 (s, 6H, H-Ar). MS (ES+) m/z: 791 (M+H)+.
EXAMPLE 10: Synthesis of 1,3,5-Tris(3,4,5-trihydroxybenzylaminomethyl)-2,4,6- triethylbenzene (19, AL 178). Following the general procedure C, the methoxy derivative 18, AL177 (170 mg, 0.21 mmol) gave 135 mg (94 %) of 19, AL178 as a white solid: mp >200 °C (dec). HPLC: tR = 2.38. 1H NMR (CD3OD, 300 MHz) δ: 0.89 (t, J = 7.3 Hz, 9H, CH3), 2.38 (q, J = 7.0 Hz, 6H, CH3-CH2), 4.12 (s, 6H, CH2-NH), 4.16 (s, 6H, CH2-NH), 6.64 (s, 6H, H-Ar). «c NMR (CD3OD, 75 MHz) δ: 16.1 (CH3), 24.9 (CH2), 43.5 (CH2), 52.6 (CH2), 111.1 (CH), 121.6 (C), 128.4 (C), 135.9 (C), 147.4 (C), 147.4 (C), 149.2 (C). MS (ES+) m/z: 664 (M+H)+. Anal. Calcd. for C42H51N3O9: C, 65.14; H, 6.83; N, 6.33. Found: C, 65.00; H, 6.63; N, 6.23.
EXAMPLE 11 : Synthesis of A/-(2-aminoethyl)-2,3,4-tris(benzyloxy)benzamide (20).
Benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP) (48 mg, 0.11 mmol, 1.2 equiv.) was added to a solution of 2,3,4-tris(benzyloxy)benzoic acid 3 (40 mg, 0.09 mmol) and triethylamine (15 μΙ, 0.11 mmol, 1.2 equiv.) in dry dichloromethane (3 ml_). After 1.5 h, the corresponding 1 ,2-diaminoetane (5.9 μΙ, 0.09 mmol) was added. The mixture was stirred at room temperature overnight; during this time a white solid was formed. The solid was filtered off and washed with dichloromethane. Evaporation of the filtrate gave a residue which was purified by CCTLC on the chromatotron using (dichlorometane/methanol/NH40H, 10:1 :0.2) as eluent. 1H NMR (CDCI3, 300 MHz) δ: 2.85 (bs, 2H, CH2-NH2), 3.35 (m, 2H, CH2-NH), 5.06 (s, 2H, CH2-Ph), 5.15 (s, 2H, CH2-Ph), 5.16 (s, 2H, CH2-Ph), 6.87 (d, J = 9.0 Hz, 1 H, H-Ar), 7.28-7.46 (m, 15H, H- Ar), 7.84 (d, J = 8.9 Hz, 1H, H-Ar), 8.22 (t, J = 5.36 Hz, 1 H, NH). EM (ES+) m/z: 483 (M+H)+. Anal. Calcd. for C42H51N3O9: C, 64.42; H, 5.24; N, 4.70. Found: C, 64.18; H, 5.20; N, 4.91. EXAMPLE 12: Synthesis of 1 ,3,5-Tris(2,3,4-tribenzyloxybenzamidoethylaminomethyl)-2,4,6- triethylbenzene (21, ECM-VI-27). A solution of 11 (50 mg, 0.11 mmol) and /V-(2-aminoethyl)- 2,3,4-tris(benzyloxy)benzamide 20 (231 mg, 0.47 mmol) was treated with triethylamine (0.065 mL, 0.47 mmol) according to the general procedure B. The residue was purified by CCTLC (dichloromethane/methanol/NH4OH, 15:1 :0.4) to give 0.11 g (57%) of 21, ECM-VI-27 as a transparent oil. HPLC: tR = 3.71. 1H NMR [(CD3)2CO, 300 MHz] δ: 1.16 (t, J = 7.2 Hz, 9H, CH3), 2.65-2.70 (m, 12 H, CH2-CH3, NH-CH2), 3.39 (q, J = 5.8 Hz, 6H, CH2-NHCO), 3.63 (s, 6H, Ar- CH2-NH), 5.06 (s, 6H, CH2-Ph), 5.11 (s, 6H, CH2-Ph), 5.16 (s, 6H, CH2-Ph), 6.88 (dd, J = 0.5, J = 9.2 Hz, 3H, H-Ar), 7.22-7.46 (m, 45H, H-Ar), 7.90 (dd, J = 0.7, J = 8.9 Hz, 3H, H-Ar), 8.03 (t, J = 5.2 Hz, 3H, NHCO). ^C-NMR (CDCI3, 75 MHz) δ: 17.14 (CH3), 22.78 (CH2), 39.69 (CH2), 47.68 (CH2), 49.52 (CH2), 70.97 (CH2), 75.77 (CH2), 109.27 (C), 120.07 (C), 126.89 (CH), 127.67 (CH), 128.29-128.96 (CH), 134.18 (C), 136.38 (C), 136.41 (C), 137.27 (C), 141.24 (C), 142.25 (C), 151.90 (C), 155.75 (C), 165.09 (C=0). EM (ES+) m/z: 1647 (M + H)+, 824 (M)2+
EXAMPLE 13: Synthesis of 1,3,5-Tris(2,3,4-trihydroxybenzamidoethylaminomethyl)-2,4,6- triethylbenzene (22, AL-184). Following the general procedure, the benzyl derivative 21, ECM- VI-27 (92 mg, 0.06 mmol) gave 45 mg (96 %) of 22, AL-184 as a white solid: mp 164-166 °C. H NMR [(CD3)OD, 300 MHz] δ: 1.19 (t, J = 7.3 Hz, 9H, CH3), 2.91 (q, 6 H, CH2-CH3), 3.50 (m, 6H, NH-CH2-CH2-NHCO), 3.81 (m, 6H, CH2-NHCO), 4.22 (s, 6H, Ar-CH2-NH), 6.39 (d, J = 8.8 Hz, 3H, H-Ar), 7.22 (d, J = 8.8 Hz, 3H, H-Ar). 13C-NMR (CDCI3. 75 MHz): 16.61 (CH3), 25.71 (ΠΗ
37.52 (CH2), 43.27 (CH2), 50.21 (CH2), 108.24 (C), 108.42 (CH), 119.95 (CH), 128.89 (C), 134.05 (C), 149.37 (C), 151.41 (C), 150.67 (C), 173.19 (C=0).
EXAMPLE 14: GENERAL SYNTHESIS METHODS AND MATERIALS FOR SYNTHESIS OF THE COMPOUNDS WITH A CENTRAL CARBOHYDRATE SCAFFOLD■ protected mono- and disaccharides.
A suspension of the corresponding sugar derivative (1 eq), the benzoic acid 2 or 3 (6.5 eq), DCC (7 eq) and DMAP (7 eq) in dry dichloromethane (70 mL) was refluxed under argon overnight. After the mixture was cooled to room temperature, the urea byproduct was filtered through celite and the filtrate was evaporated. The resulting residue was purified by column chromatography. For disaccharides (maltose and trehalose) 10 eq of DCC and DMAP was used.
EXAMPLE 15: Synthesis of or- and j3-D-Mannopyranose pentakis[2,3,4- tris(phenylmethoxy)benzoate] (35 and 36 PCB248). Following the general procedure D- mannose (50 mg, 0.28 mmol) was treated with 3 (713 mg, 1.62 mmoles, 6 eq), DCC (33,4 mg, 1.62 mmol, 6 eq) and DMAP (198 mg, 1.62 mmol, 6 eq) to give a residue that was purified by column chromatography on silica gel using a mixture of toluene/ethyl acetate (96:4). From the fastest running fraction 203 mg (33%) of 35 PCB248 alpha {a anomer) was obtained as a white solid. 1H NMR (CDCI3, 300 MHz) δ: 4.41-4.50 (m, 2H, H-6), 4.65 (bs, 1 H, H-5), 4.85-5.29 (m, 30H, CH2-Ph), 5.91 (m, 1 H, H-2), 6.09 (dd, J = 3.0 Hz, J = 10.2 Hz, 1 H, H-3), 6.12 (m, 1 H, H-4), 6.40 (d, J = 9.1 Hz, 1 H, H-Ar), 6.55-6.61 (m, 4H, H-Ar and H-1), 6.84 (d, J = 8.9 Hz, 1 H, H-Ar), 6.90 (d, J = 9.1 Hz, 1 H, H-Ar), 7.19-7.70 (m, 77H, H-Ar), 7.80 (d, J = 8.9 Hz, 1 H, H-Ar), 7.87 (d, J = 8.9 Hz, 1 H, H-Ar).
From the slowest running fraction 124.2 mg (20%) of 36 PCB248 beta (β anomer) was obtained as a white solid. Mp 188-190 °C. (CDCI3, 300 MHz) <5: 4.49-4.68 (m, 3H, H-5, H-6), 4.82-5.17 (m, 30H, CH2-Ph), 5.71 (dd, J = 3.0 Hz, J = 10.0 Hz, 1 H, H-3), 6.03 (t, J = 9.9 Hz, 1 H, H-4), 6.09 (d, J = 3.2 Hz, 1 H, H-2), 6.23 (s, 1 H, H-1), 6.40 (dd, J = 2.3 Hz, J = 9.0 Hz, H-Ar), 6.55 (d, J = 8.9 Hz, 1 H, H-Ar), 6.62 (dd, J = 1.2 Hz, J = 9.0 Hz, 2H, H-Ar), 7.20-7.68 (m, 78H, H-Ar), 7.80 (d, J = 8.9 Hz, 1 H, H-Ar), 7.86 (d, J = 8.9 Hz, 1 H, H-Ar).
EXAMPLE 16: Synthesis of a-D-Mannopyranose pentakis(2,3,4-trihydroxybenzoate) (37, AL-173). Following the general procedure C the benzyl protected derivative 35 gave 63.6 mg (78% yield) of the deprotected a anomer 37 AL173 as a white solid. Mp 158-160 °C. [a]24 D + 4.0 0 (c 0.15, acetone). HPLC: t = 3.58 min. 1H NMR (CD3OD, 400 MHz) δ: 4.52-4.68 (m, 3H
and H-6), 5.93 (dd, J = 2.0 Hz, J = 3.2 Hz, H-2), 5.98 (dd, J = 3.2 Hz, J = 10.2 Hz, 1 H, H-3), 6.15 (t, J = 10.1 Hz, 1 H, H-4), 6.20 (d, J = 8.9 Hz, 1 H, H-Ar), 6.35 (d, J = 8.9 Hz, 1 H, H-Ar), 6.38 (d, J = 8.8 Hz, 1 H, H-Ar), 6.45 (d, J = 8.8 Hz, 1 H, H-Ar), 6.56 (d, J = 8.8 Hz, 1 H, H-Ar), 6.57 (d, J = 1.9 Hz, 1 H, H-1), 6.95 (d, J = 8.9 Hz, 1 H, H-Ar), 7.32 (d, J = 8.9 Hz, 1 H, H-Ar), 7.39 (d, J = 8.8 Hz, 1 H, H-Ar), 7.46 (d, J = 8.8 Hz, 1 H, H-Ar), 7.57 (d, J = 8.8 Hz, 1 H, H-Ar). «C NMR (CD3OD, 100 MHz) δ: 62.69 (CH2), 66.60 (CH), 70.27 (CH), 71.35 (CH), 72.44 (CH), 92.65 (CH), 105.00- 105.84 (C), 108.88-109.31 (CH), 122.39-122.85 (CH), 133.51-133.95 (C) 152.25-153.84 (CH and C), 168.91-170.88 (C=0). ). EM (ES+) m/z: 941 (M+H)+, 771. Anal. Calcd. for CesHs^s: C, 52.35; H, 3.43. Found: C, 52.08; H, 3.71.
EXAMPLE 17: Synthesis of /3-D-Mannopyranose pentakis(2,3,4-trihydroxybenzoate) (38, AL-174). Following the general procedure C the benzyl protected derivative 36 gave 33.2 mg (80 %) of the deprotected β anomer 38 AL174 as a white solid. Mp 160-162 °C. [a]24 D +79.9° (c 0.154, acetone). HPLC: tR(S) = 3.50 min. 1H NMR (CD3OD, 400 MHz) δ: 4.49 (dt, J = 3.0 Hz, J = 9.8 Hz, 1 H, H-5), 4.65 (ddd, J = 3.0 Hz, J = 12.5 Hz, J = 38.9 Hz, 2H, H-6), 5.92 (dd, J = 3.2 Hz, J = 9.9 Hz, 1 H, H-3), 6.05 (t, J = 9.9 Hz, 1 H, H-4), 6.08 (dd, 1 H, J = 1.0 Hz, J = 3.2 Hz, H-2), 6.22 (d, J = 8.9 Hz, H-Ar), 6.25 (d, J = 8.9 Hz, 1 H, H-Ar), 6.37 (d, J = 8.9 Hz, 1 H, H-Ar), 6.41 (d, J = 8.8 Hz, 1 H, H-Ar), 6.46 (d, J = 8.8 Hz, 1 H, H-Ar), 6.53 (d, J = 1.1 Hz, 1 H, H-1), 6.99 (d, J = 9.8 Hz, 1 H, H-Ar), 7.01 (d, J = 9.8 Hz, 1 H, H-Ar), 7.31 (d, J = 7.3 Hz, 1 H, H-Ar), 7.48 (d, J = 8.3 Hz, 1 H, H-Ar), 7.49 (d, J = 8.3 Hz, 1 H, H-Ar). "C NMR (CD3OD, 100 MHz) δ: 61.81 (CH2), 66.08 (CH), 69.55 (CH), 71.30 (CH), 72.72 (CH), 90.96 (CH), 103.63-104.67 (C), 107.68-108.09 (CH), 121.26-121.47 (CH), 132.34-132.65 (C), 151.10-152.43 (CH and C), 167.77-169.72 (C=0). EM (ES+) m/z: 941 (M+H)+, 771. Anal. Calcd. for C68H54043: C, 52.35; H, 3.43. Found: C, 52.07; H, 3.71.
EXAMPLE 18: Synthesis of 0-D-Maltose octakis[3,4,5-tris(phenylmethoxy)benzoate] (39 AL34). Following the general procedure monohydrated Maltose (50 mg, 0.14 mmol) was treated with 210 (607 mg, 1.38 mmoles, 10 eq), DCC (284 mg, 1.38 mmol, 10 eq) and DMAP (169 mg, 1.38 mmol, 10 eq) to give a residue that was purified by column chromatography on silica gel using a mixture of dichloromethane/toluene/ethyl acetate 75:25:1 to give 440 mg (86%) of 39 AL34 as a white solid. Mp 59-61 °C. [a]24 D -12,3° (c 0.153, CH2CI2). HPLC: tR(S) = 3.78. 1H NMR (CDC , 300 MHz) δ: 4.20-4.57 (m, 7H, H-4, H-5, H-6 and H-6'), 4.62-5.08 (m, 48H, CH2-Ph), 5.36 (dd, J = 3.7 Hz, J = 10.7 Hz, 1 H, H-2'), 5.50-5.60 (m, 2H, H-1' and H-2), 5.69 (t, J = 9.9 Hz, 1 H,
Η-4'), 6.00 (t, J = 9.7 Hz, 1H, H-3), 6.09-6.17 (m, 2H, H-1 and H-3'), 6.82-7.38 (m, 136H, CH-Ar). EM (ES+) m/z: 3744 (M+Na)+.
EXAMPLE 19: Synthesis of /J-D-Maltose octakis(3,4,5-trihydroxybenzoate) (40, AL175).
Following the general procedure C the benzyl protected derivative 39 (125 mg, 0.034 mmol) gave
44.3 mg (82% yield) of the deprotected a anomer 40 as a white solid. Mp >190 °C (dec).[a]24D +38.3° (c 0.152, acetone). HPLC: tR(S) = 2.84 min. 1H NMR (CD3OD, 400 MHz) δ: 4.12 (bs, 2H, H-6'), 4.21 (dt, J = 2.0 Hz, J = 10.0 Hz, 1 H, H-5'), 4.38 (dt, J = 2.7 Hz, J = 9.7 Hz, 1 H, H-5), 4.59 (t, J = 9.3 Hz, 1 H, H-4), 4.74 (bs, 2H, H-6), 5.13 (dd, J = 3.9 Hz, J = 10.5 Hz, 1 H, H-2'), 5.43 (dd, J = 8.4 Hz, J = 9.8 Hz, 1 H, H-2), 5.55 (t, J = 9.8 Hz, 1 H, H-4'), 5.78 (d, J = 9.8 Hz, 1 H, H-1 '), 5.82- 5.91 (m, 2H, H-3 and H-3'), 6.17 (d, J = 8.3, 1 H, H-1), 6.80 (s, 2H, CH-Ar), 6.84 (s, 2H, CH-Ar), 6.87 (s, 2H, CH-Ar), 6.89 (s, 2H, CH-Ar), 6.95 (s, 2H, CH-Ar), 7.02 (s, 2H, CH-Ar), 7.07 (s, 2H, CH-Ar), 7.20 (s, 2H, CH-Ar). 13C NMR (CD3OD, 100 MHz) δ: 62.56, 63.40, 64.00, 69.49, 70.64, 71.14, 71.78, 72.71 , 72.77, 74.75, 76.30, 93.60, 97.63, 110.29-110.98 (CH and C), 119.77-121.04 (C), 139.96-140.752 (C), 146.03-146.75 (CH and C), 166.26-168.24 (C=0). EM (ES+) m/z: 1559 (M+H)+. Anal. Calcd. for C68H54O43: C, 52.38; H, 3.49. Found: C, 52.02; H, 4.65.
EXAMPLE 20: Synthesis of α,α-Trehalose octakis[3,4,5-tris(phenylmethoxy)benzoate] (41, AL35). Following the general procedure dehydrated α,α-Trehalose (50 mg, 0.14 mmol) was treated with 2 (581 mg, 1.32 mmoles, 10 eq), DCC (272.3 mg, 1.32 mmol, 10 eq) and DMAP (161.26 mg, 1.32 mmol, 10 eq) to give a residue that was purified by column chromatography on silica gel using a mixture of dichloromethane/toluene/ethyl acetate 75:25:1 to give 398 mg (81%) of 41 AL35 as a white solid. Mp 83-85 °C. [a]24 D +79.7° (c 0.156, CH2CI2). HPLC: tR = 3.75 min. Ή NMR (CDCI3, 300 MHz]) δ: 4.27-4.32 (m, 4H, H-6), 4.41-4.47 (m, 2H, H-5), 4.55-5.15 (m, 48H, CH2-Ph), 5.21 (dd, J = 3.8 Hz, J = 10.1 , 2H, H-4), 5.62 (t, J = 9.9 Hz, 2H, H-2), 3.97 (d, J = 4.0 Hz, 2H, H-1), 6.38 (t, J = 10.0 Hz, 2H, H-3), 6.89-7.40 (m, 136H, Η Γ). EM (ES+) m/z: 3744 (M+Na)+.
EXAMPLE 21 : Synthesis of α,α-Trehalose octakis (3,4,5-trihydroxybenzoate) 42 (AL166).
Following the general procedure C the benzyl protected derivative 41 (200 mg, 0.053 mmol) gave 72 mg (87 % yield) of 42 as a white solid. Mp >200 °C (dec). [a]24 D +19.7° (c 0.155, acetone). HPLC: ttw = 9.18 min. 1H NMR (CD3OD, 300 MHz) δ: 3.63 (d, J = 11.7 Hz, 2H, H-6), 3.93 (d, J =
12.4 Hz, 2H, H-6), 4.22 (d, J = 10.3 Hz, 2H, H-5), 5.26 (dd, J = 4.0 Hz, J = 10.3 Hz, 2H, " " "
5.60 (t, J = 10.1 Hz, 2H, H-2), 5.70 (d, J= 3.7 Hz, 2H, H-1), 6.02 (t, J = 10.1 Hz, 2H, H-3), 6.90 (s, 4H, H-Ar), 6.98 (s, 4H, H-Ar), 7.05 (s, 4H, H-Ar), 7.20 (s, 4H, H-Ar). EM (ES+) m/z: 1559 (M+H)+. Anal. Calcd. for C68H54O43: C, 52.38; H, 3.49. Found: C, 52.61 ; H, 3.64. EXAMPLE 22: Surface Plasmon Resonance Experiments for evaluation of the binding of the compounds of the invention to gp120.
Recombinant gp120 protein from the HIV-1 NIB strain (ImmunoDiagnostics Inc., Woburn, MA) (produced in cell cultures of Chinese hamster ovaries) was covalently immobilized on the carboxymethylated dextran matrix of a CM5 sensor chip in 10 mM sodium acetate, pH 4.0 using standard amine coupling chemistry up to a final density of 1770 RUs (15 fmol of gp120). A reference flow cell was used as a control for non-specific binding and refractive index changes. All interaction studies were performed at 25 °C on a Biacore T100 instrument (GE Healthcare, Uppsala, Sweden). The compounds were diluted in HBS-P (10 mM HEPES, 150 mM NaCI and 0,05% surfactant P20; pH 7.4) supplemented with 5% dimethyl sulfoxide (DMSO, Merck) and 10 mM Ca2+ at a concentration of 50 μΜ. Samples were injected for 3 minutes at a flow rate of 30 μΙ/min followed by a dissociation phase of 5 minutes. A DMSO concentration series was included to eliminate the contribution of DMSO to the measured response. The CM5 sensor chip surface was regenerated with an injection of 50 mM NaOH for 5 seconds at a flow rate of ΙΟΟμΙ/min. The compounds that showed significant binding to gp120 are shown in table 1. The affinity of the compounds is evaluated through the response units obtained from the binding of the compounds to the chip both at the end of the injection phase and after 50s of dissociation.
Table 1 : Response units (RUs) of the binding of the synthesized compounds to gp120 on a CM5 sensor chip at the end of the injection phase and after 50 sec of dissociation.
The same general methods were used as described for other compounds. For HPLC, acetonitrile was used as mobile phase A with 0.08% of formic acid, and water was used as mobile phase B with 0.1% of formic acid at a flow rate of 1 ml/min on a SunFire Os (4.6 x 50 mm, 3.5 μΐτι) column with a A:B, 0-100% of A.
Synthesis of 1-aminomethyl-3,5-Bis(2,3,4-tribenzyloxybenzoylaminomethyl)-2,4,6-triet^
(103). To a solution containing the benzyl protected trihydroxybenzoic acid 102 (200 mg, 0.45 mmol) (Takashi, N., at al. JP 08143525 A 19960604) and PyBOP (263 mg, 0.50 mmol) in dichloromethane (3 ml_), EfeN (70 μΐ_, 0.50 mmol) was added. The mixture was stirred at room temperature for 1 h, and added dropwise to a suspension of the trisamine derivative 101 (63 mg, 0.25 mmol)(Walsdorff, C, et al. J. Chem.Res (M) 1996, 1601-1609; Wallace, K. J., et al. Synthesis 2005, 12, 2080-2083). in dichloromethane (1 ml_). The mixture was stirred at room temperature for 2 h. Dichloromethane (50 mL) was added and the reaction mixture was washed with brine (20 mL). The organic phase was dried (anhydrous MgS04), filtered, and evaporated to dryness. The residue was purified by CCTLC on the Chromatotron using (dichloromethane/methanol, 20:1) as eluent to afford 114 mg (41%) of 103 as a pale yellow solid. HPLC: tR = 4.75 min. Ή NMR (CDCI3, 300 MHz) δ: 0.90-1.28 (m, 9H, CH3CH2), 2.44 (m, 2H, CH3CH2), 2.66 (m, 4H, CH3CH2), 3.95 (m, 2H, CH2NH2), 4.54-4.60 (m, 4H, CH2NH), 4.85-4.96 (m, 8H, CH2Ph), 5.15 (m, 4H, CH2Ph), 6.79-7.41 (m, 32, H-Ar), 7.87 (m, 2H, NH), 7.95 (m, 2H, H- Ar). EM (ES+) m/z: 1094 (M+1).+
Synthesis of 1-A/-I (13'-Fmoc-amino-4',7',10'-trioxatridecyl)succinamoyl1aminomethyl-3,5- Bis(2,3,4-tribenzyloxybenzoylaminomethyl)-2,4,6-triethylbenzene (105). To a solution of 103 (233 mg, 0.21 mmol) and PyBOP (222 mg, 0.43 mmol) in dichloromethane (2 mL), N-Fmoc-N'- succinyl-4,7,10-trioxa-1 ,13-tridecanediamine 104 (232 mg, 0.43 mmol) (Reina, J.J., et al. Bioconjugate Chem. 2007, 18, 963-969) and EtjN (60 μΐ, 0.43 mmol) were added at 0 °C. The reaction mixture was stirred at 0 °C for 1h. Dichloromethane (50 mL) was added and the reaction mixture was washed with brine (20 mL). The organic phase was dried (anhydrous MgSC ), filtered, and evaporated to dryness. The residue was purified by CCTLC on the Chromatotron using (dichloromethane/methanol, 20:1) as eluent to afford 326 mg (95%) of 105 as a pale yellow solid. HPLC: tR = 7.20 min. Ή NMR (CDCI3, 300 MHz) δ: 1.03-1.15 (m, 9H, CH3CH2), 1.67-1.78 (m, 4H, CH2), 2.23 (m, 2H, CH2CH2), 2.38-2.50 (m, 4H, CH3CH2, Ctf2CH2), 2.66 (m, 4H, CH3CH2), 3.48-3.63 (m, 12H, CH2), 3.35 (m, 4H, CH2), 4.21 (m, 2H, Ctf2NH), 4.32-4.41 (m, 3H, Fmoc), 4.60 (m, 4H, CH2NH), 4.88 (s, 4H, CH2Ph), 4.91 (s, 4H, CH2Ph), 5.14 (s, 4H, CH2Ph), 7.05 (m, 2H, H-Ar), 7.14-7.41 (m, 34H, H-Ar, Fmoc), 7.59 (m, 2H, Fmoc), 7.75 (m, 2H, Fmoc), 7.89 (m, 2H, NH), 7.96 (d, J = 9 Hz, 2H, H-Ar). EM (ES+) m/z: 1620 (M+1).+
Synthesis of 1-A/-[(13'- amino-4',7',10'-trioxatridecyl)succinamoyl1aminomethyl-3,5-Bis(2,3,4- tribenzyloxybenzoylaminomethyl)-2,4,6-triethylbenzene (106). A solution of 105 (325 mg,
mmol) in dichloromethane (2 mL) was treated with piperidine (0.5 mL). The reaction mixture was stirred at room temperature for 1 h, and the volatiles were removed. The residue was purified by CCTLC on the Chromatotron using (dichloromethane/methanol, 10:1) as eluent to afford 217 mg (77%) of 106 as a pale yellow solid. HPLC: tR = 4.56 min. Ή NMR (DMSO-d6, 300 MHz) δ: 0.92- 1.03 (m, 9H, CH3CH2), 1.57 (m, 2H, CH2), 1.77 (m, 2H, CH2), 2.25 (m, 4H, CH2CH2), 2.50-2.59 (m, 2H, CH2NH2), 2.81 (m, 2H, CH3CH2), 3.03 (m, 4H, CH3CH2), 3.37-3.52 (m, 14H, CH2), 4.21 (m, 2H, CH2NH), 4.45 (m, 4H, CH2NH), 4.89 (s, 4H, CH2Ph), 4.91 (s, 4H, CH2Ph), 5.19 (s, 4H, Ctf2Ph), 7.03-7.57 (m, 34H, H-Ar), 7.89 (m, 4H, NH). EM (ES+) m/z: 1397 (M+1).+ Synthesis of 1-A/-I (13 - amino-4',7',10'-trioxatridecyl)succinamoyl1aminomethyl-3,5-Bis(2,3,4- trihvdroxybenzoylaminomethyl)-2,4,6-triethylbenzene (107, AL202). A solution of 106 (129 mg, 0.09 mmol) in THF/methanol (1 :1) (8 mL) containing 30 wt% of Pd/C (10%) was hydrogenated at 2.85 atm (42 psi) at 30 °C overnight. The Pd/C was filtered through Whatman® filter paper 42 and the solvent was removed under reduced pressure to afford 70 mg (89%) of 107 (AL202) as a pale yellow solid. HPLC: tR = 2.96 min. 1H NMR (DMSO-d6, 300 MHz) <5: 1.17-1.21 (m, 9H, CH3CH2), 1.71 (m, 2H, CH2), 1.90 (m, 2H, CH2), 2.47 (s, 2H, CH2CH2), 2.79-2.91 (m, 6H, CH2CH3), 3.07 (m, 2H, CH2), 3.20 (m, 2H, CH2), 3.44-3.66 (m, 12H, CH2), 4.45 (m, 2H, CH2NH), 4.66 (s, 4H, CH2NH), 6.34 (d, J = 8.8 Hz, 2H, H-Ar), 7.21 (d, J = 8.8 Hz, 2H, H-Ar). EM (ES+) m/z 856 (M+1).+
Claims
1. A compound with a structure according to formula (A), (B), (C), (D), or (E)
(B)
- each n is independently selected from 1 and 2;
- L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, and heteroalkynylene can be unsubstituted or substituted with =0, =S; OH; SH; and NH2;
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
heteroalkynylene; and 1 ,4-piperazinylene; - each of R1a, R1b and R1c is independently selected from methyl; and ethyl;
- each of R, R3, R4 and R5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; dihydroxybenzyl; trihydroxybenzyl; dimethoxyphenyl; trimethoxyphenyl; dimethoxybenzyl;
trimethoxybenzyl, and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3 and wherein each Y is independently selected from hydrogen, methyl or benzyl; or
- one of X-R3, X-R4 or X-R5 is a linking moiety selected from Ci.iooheteroalkyl; Ci-iooheteroalkenyl; and Ci-iooheteroalkynyl, wherein said Ci.iooheteroalkyl, Ci-iooheteroalkenyl and Ci- looheteroalkynyl can be unsubstituted or substituted with one or more =0, =S; OH; -SH; and NH2; or
wherein Q is selected from H2N(CH2)30(CH2)20(CH2)3NH-, H2N(CH2)20(CH2)2NH-, H2N(CH2)30 (CH2)3NH-, H2N(CH2)(CH2)i-i2(CH2)NH-, H2 (CH2)30(CH2)20(CH2)3NH-, H2N(CH2)2NH(CH2)2NH- , H2N(CH2)3NH(CH2)i NH-„ H2N(CH2)3N(CH3)(CH2)3NH-, H2N(CH2)2NH(CH2)2-3(CH2)2NH-, H2N(CH2)3NH(CH2)2-4(CH2)2NH(CH2)2NH -, H2N(CH2)3N(CH2 CH2)NH(CH2)3NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)3NH(CH2)CH=CH(CH2)NH(CH2)3NH -;
with the proviso that said compound is not
- 1 ,3,5-Tris(2,3-dimethoxybenzamidomethyl)-2,4,6-triethylbenzene (9, PCB311);
- 1 ,3,5-Tris(2,3-dihydroxybenzamidomethyl)-2,4,6-triethylbenzene (10, AL180);
- a- or jS-D-Glucopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (23, AL-18);
- a- or jS-D-Glucopyranose pentakis[3,5-bis(phenylmethoxy)benzoate];
- a- or jS-D-Glucopyranose pentakis[3,4-bis(phenylmethoxy)benzoate];
- σ or /3-D-Glucitol 1 ,5-anhydro-pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j6-D-Galactopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (25, AL-29 and AL-30);
- a- or jS-D-Allopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j8-D-Mannopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate](29, 30 and 31);
- a- or -D-Xylopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or jS-L-Arabinopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or -D-Ribopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate] (27, AL36);
- a- or j8-D-Luxopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j8-D-Glucopyranose pentakis[3,5-dihydroxy)benzoate]; - o- or -D-Glucopyranose pentakis[3,4-dihydroxybenzoate];
- D-Galactopyranose pentakis[3,4-dihydroxybenzoate];
- D-Arabinopyranose tetrakis[3,4-dihydroxybenzoate];
- D-Mannopyranose pentakis[3,4-dihydroxybenzoate];
- σ or β-D-Glucitol 1 ,5-anhydro-pentakis[3,4,5-trihydroxybenzoate];
- or- or j8-D-Glucopyranose pentakis[3,4,5-trihydroxybenzoate] (24, AL-19);
- a- or j8-D-Galactopyranose pentakis[3,4,5-trihydroxybenzoate] (26, AL-33);
- or- or jS-D-Allopyranose pentakis[3,4,5-trihydroxybenzoate];
- a- or -D-Mannopyranose pentakis[3,4,5-trihydroxybenzoate](32, AL51a and 33, AL51 b);
- a- or β-D-Xylopyranose tetrakis[3,4,5-trihydroxybenzoate];
- or- or j8-L-Arabinopyranose tetrakis[3,4,5-trihydroxybenzoate];
- σ- or β-D-Ribopyranose tetrakis[3,4,5-trihydroxybenzoate] (28, AL50);
- a- or j8-D-Luxopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or β-D-Glucopyranose pentakis[3,4,5-trimethoxybenzoate];
- σ- or -D-Mannopyranose pentakis[3,4,5-trimethoxybenzoate]
-]8-D-Glucopyranose1-[4,5-dihydroxy-2-(phenylmethoxy)benzoate]2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate];
- β -D-Glucopyranose, 1-[3,4-dihydroxy-5-(phenylmethoxy)benzoate] 2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate].
2. The compounds according to claim 1 , wherein n is 1.
3. The compounds according to claims 1 and 2, wherein each X is independently selected from - 0-; -NH-; -NH-(CH2)-; -NH-(CH2)2-NH-; -0-(CH2)i-i8-0-; -0-(CH2)2-0-(CH2)2-0-; -NH-(CH2)3-0- (CH2)2-0-(CH2)2-0-(CH2)3-NH-; -NH-(CH2-CH20)m-CH2-CH2-NH-; -NH-CH2-CH2-CH2-(0-CH2-
CH2)i-CH2-NH-; — N^¾N_ ; -NH-CH2-Z-CH2-NH-, wherein each of m and I are
independently selected from 0, 1 , 2, 3, 4 and 5, and wherein Z is independently selected from - (CH2)i-i2-; -CH2-NH-CH2-; -CH2-CH2-NH-(CH2)i-3-; -CH2-CH2-N(-CH3)-CH2-CH2-; -CH2-NH-(CH2)2- 3-NH-CH2-; -CH2-CH2-NH-(CH2)2.4-NH-CH2-CH2-; -CH2-CH2-N(-CH2-CH2)2-N-CH2-CH2-; -CH2-NH- (CH2)2-NH-(CH2)2-NH-CH2-; -CH2-NH-(CH2)2-NH-(CH2)2-NH-(CH2)2-NH-CH2-; -CH2-CH2-NH-CH2- CH=CH-CH2-NH-(CH2)2-.
4. The compounds according to claims 1 to 3, wherein each of R1a, R1b and R1c is ethyl.
5. The compounds according to claims 1 to 4, wherein the compounds have a structure of formula (A) or (B) wherein each X is independently selected from -NH-; or -NH-(CH2)2-NH-; each of R3, R4 and R5 is independently selected from a structure with formula (I), k is 3; (OY) is in the 3,4,5-position or the 2,3,4-position; and each Y is hydrogen.
6. The compounds according to claims 1 to 5, wherein the compounds are selected from the list of:
- 1 ,3,5-Tris(3,4,5-tribenzyloxybenzamidomethyl)-2,4,6-triethylbenzene (5, PCB245);
- 1 ,3,5-Tris(3,4,5-trihydroxybenzamidomethyl)-2,4,6-triethylbenzene (6, AL 113);
- 1 ,3,5-Tris(2,3,4-tribenzyloxybenzamidomethyl)-2,4,6-triethylbenzene (7, AL 168);
- 1 )3)5-Tris(2,3,4-trihydroxybenzamidomethyl)-2,4,6-triethylbenzene (8, AL 170);
- 1 ,3,5-Tris(3,4-dimethoxyphenylaminomethyl)-2,4,6-triethylbenzene (15, AL-125);
- 1 ,3,5-Tris(3,4-dihydroxyphenylaminomethyl)-2,4,6-triethylbenzene (16, AL-172);
- 1 ,3,5-Tris(3,4,5-trimethoxyphenylaminomethyl)-2,4,6-triethylbenzene (17, AL 126);
- 1 ,3,5-Tris(3,4,5-trimethoxybenzylaminomethyl)-2,4,6-triethylbenzene (18, AL 177);
- 1 ,3,5-Tris(3,4,5-trihydroxybenzylaminomethyl)-2,4,6-triethylbenzene (19, AL 178);
- 1 ,3,5-Tris(2,3,4-tribenzyloxybenzamidoethylaminomethyl)-2,4,6-triethylbenzene (21 , ECM-VI- 27);
- 1 ,3,5-Tris(2,3,4-trihydroxybenzamidoethylaminomethyl)-2,4,6-triethylbenzene (22, AL-184);
- a- or -D-Mannopyranose pentakis[2,3,4-tris(phenylmethoxy)benzoate] (34,35 and 36 PCB248);
- a- D-Mannopyranose pentakis(2,3,4-trihydroxybenzoate) (37, AL-173);
- β- D-Mannopyranose pentakis(2,3,4-trihydroxybenzoate) (38, AL-174);
- β- D-Maltose octakis[3,4,5-tris(phenylmethoxy)benzoate] (39, AL34);
- β- D-Maltose octakis(3,4,5-trihydroxybenzoate) (40, AL175);
- a, a-Trehalose octakis[3,4,5-tris(phenylmethoxy)benzoate] (41 , AL35); and
- a, a-Trehalose octakis (3,4,5-trihydroxybenzoate) 42 (AL166) ; and
- 1-A/-[ (13'- amino-4',7',10'-trioxatridecyl)succinamoyl]aminomethyl-3,5-Bis(2,3,4- trihydroxybenzoylaminomethyl)-2,4,6-triethylbenzene.
7. A compound for targeting HIV gp120
wherein the compound has a structure according to formula (A), (B), (C), (D), or (E)
- each n is independently selected from 1 and 2;
- L is selected from alkylene; alkenylene; alkynylene; heteroalkylene; heteroalkenylene; and heteroalkynylene; wherein said alkylene, alkenylene, alkynylene, heteroalkylene, heteroalkenylene, and heteroalkynylene can be unsubstituted or substituted with =0, =S; OH; SH; and NH2;
- each X is independently selected from -0-; -NH-; heteroalkylene; heteroalkenylene;
heteroalkynylene; and 1 ,4-piperazinylene;
- each of R1a, R1b and R1c is independently selected from methyl; and ethyl;
- each of R, R3, R4 and R5 is independently selected from dihydroxyphenyl; trihydroxyphenyl; dihydroxybenzyl; trihydroxybenzyl; dimethoxyphenyl; trimethoxyphenyl; dimethoxybenzyl; trimethoxybenzyl, and the structures with formula (I), (II), (III) or (IV), wherein k is selected from 2 and 3 and wherein each Y is independently selected from hydrogen, methyl or benzyl; or
- one of X-R3, X-R4 or X-R5 is a linking moiety selected from Ci-iooheteroalkyl; Ci-iooheteroalkenyl; and Ci-iooheteroalkynyl, wherein said Ci-iooheteroalkyl, Ci-iooheteroalkenyl and Ci- looheteroalkynyl can be unsubstituted or substituted with one or more =0, =S; OH; -SH; and NH2; or
- one of X-R3, X-R4 or X-R5 is a linking moiety selected from
wherein Q is selected from H2N(CH2)30(CH2)20(CH2)3NH-, H2N(CH2)20(CH2)2NH-, H2N(CH2)30 (CH2)3NH-, H2N(CH2)(CH2)i-i2(CH2)NH-, H2N(CH2)30(CH2)20(CH2)3NH-, H2N(CH2)2NH(CH2)2NH- , H2N(CH2)3NH(CH2)i-4 NH-„ H2N(CH2)3N(CH3)(CH2)3NH-, H2N(CH2)2NH(CH2)2-3(CH2)2NH-, H2N(CH2)3NH(CH2)2 (CH2)2NH(CH2)2NH -, H2N(CH2)3N(CH2 CH2)NH(CH2)3NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH(CH2)2NH -,
H2N(CH2)3NH(CH2)CH=CH(CH2)NH(CH2)3NH -.
8. The compound according to claim 7, wherein said compound is selected from the list of:
- 1 ,3,5-Tris(2,3-dihydroxybenzamidomethyl)-2,4,6-triethylbenzene (9, PCB311);
- 1 ,3,5-Tris(2,3-dihydroxybenzamidomethyl)-2,4,6-triethylbenzene (10, AL180);
- 1 ,3,5-Tris(2,3-dimethoxybenzamidomethyl)-2,4,6-triethylbenzene;
- 1 ,3,5-Tris(2,3-dimethoxybenzamidomethyl)-2,4,6-triethylbenzene;
- a- or β-D-Glucopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (23, AL-18);
- a- or jS-D-Glucopyranose pentakis[3,5-bis(phenylmethoxy)benzoate];
- o- or -D-Glucopyranose pentakis[3,4-bis(phenylmethoxy)benzoate];
- σ or β-D-Glucitol 1 ,5-anhydro-pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- σ- or j3-D-Galactopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate] (25, AL-29 and AL-30);
- a- or j8-D-Allopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or -D-Mannopyranose pentakis[3,4,5-tris(phenylmethoxy)benzoate](29, 30 and 31);
- a- or j8-D-Xylopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j8-L-Arabinopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j6-D-Ribopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate] (27, AL36);
- a- or jS-D-Luxopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or j8-D-Glucopyranose pentakis[3,5-dihydroxy)benzoate]; - a- or j8-D-Glucopyranose pentakis[3,4-dihydroxy)benzoate];
- D-Galactopyranose pentakis[3,4-dihydroxy)benzoate];
- D-Arabinopyranose tetrakis[3,4-dihydroxy)benzoate];
- D-Mannopyranose pentakis[3,4-dihydroxy)benzoate];
- a or jS-D-Glucitol 1 ,5-anhydro-pentakis[3,4,5-trihydroxybenzoate];
- a- or β-D-Glucopyranose pentakis[3,4,5-trihydroxybenzoate] (24, AL-19);
- a- or ]8-D-Galactopyranose pentakis[3,4,5-trihydroxybenzoate] (26, AL-33);
- a- or jS-D-Allopyranose pentakis[3,4,5-trihydroxybenzoate];
- o- or jS-D-Mannopyranose pentakis[3,4,5-trihydroxybenzoate](32, AL51a and 33, AL51b); - a- or jS-D-Xylopyranose tetrakis[3,4,5-tris(phenylmethoxy)benzoate];
- a- or jS-L-Arabinopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or -D-Ribopyranose tetrakis[3,4,5-trihydroxybenzoate] (28, AL50);
- a- or jS-D-Luxopyranose tetrakis[3,4,5-trihydroxybenzoate];
- a- or 3-D-Glucopyranose pentakis[3,4,5-trimethoxybenzoate];
- or- or j3-D-Mannopyranose pentakis[3,4,5-trimethoxybenzoate]
-jS-D-Glucopyranose1-[4,5-dihydroxy-2-(phenylmethoxy)benzoate]2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate];
- β -D-Glucopyranose, 1-[3,4-dihydroxy-5-(phenylmethoxy)benzoate] 2,3,4,6-tetrakis[3,4,5- tris(phenylmethoxy)benzoate].
9. A pharmaceutical composition comprising a compound as claimed in claim 1 to 8 linked to an HIV inhibitor or antibody-recruiting therapeutic.
10. A process for preparing a pharmaceutical composition as claimed in claim 9 wherein the compound as claimed in claim 1 to 8 is intimately mixed with one or more pharmaceutical excipients or pharmaceutically acceptable carriers.
11. The composition of claim 9 for the treatment of an HIV infection.
12. Use of the composition as claimed in claim 7 and 8 for the manufacture of a medicament for the treatment of an HIV infection.
13. A method for (a) binding or purifying (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus, or (3) gp120 comprising viruses or for (b) separating (1) the protein gp120 or fragments thereof, (2) cells infected with a gp120 comprising virus or (3) gp120 comprising viruses, from other proteins, cells or viruses which do not comprise gp120, wherein said method comprises the use of a compound with a structure according to claims 1 to 8.
14. The method according to claim 13, wherein the compound with a structure according to claims 1 to 8 is covalently coupled to a polymer.
Applications Claiming Priority (8)
| Application Number | Priority Date | Filing Date | Title |
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| GB1000742.5 | 2010-01-18 | ||
| GB1000746.6 | 2010-01-18 | ||
| GBGB1000746.6A GB201000746D0 (en) | 2010-01-18 | 2010-01-18 | Gp 120 binding compounds |
| GBGB1000742.5A GB201000742D0 (en) | 2010-01-18 | 2010-01-18 | Gp120 binding compounds and their use |
| GBGB1001404.1A GB201001404D0 (en) | 2010-01-29 | 2010-01-29 | Gp120 Binding compounds and their use |
| GBGB1001405.8A GB201001405D0 (en) | 2010-01-29 | 2010-01-29 | Gp120 binding compounds |
| GB1001404.1 | 2010-01-29 | ||
| GB1001405.8 | 2010-01-29 |
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| WO2020104687A1 (en) * | 2018-11-24 | 2020-05-28 | Momentive Performance Materials Gmbh | Use of polyhydroxyaromatic compounds for the treatment of fibrous amino acid based substrates |
| WO2021155864A1 (en) * | 2020-02-08 | 2021-08-12 | Syneurx International (Taiwan) Corp. | Compounds and pharmaceutical uses thereof |
| CN114685574A (en) * | 2020-12-31 | 2022-07-01 | 中国科学院上海药物研究所 | Polyphenol compound and preparation method and application thereof |
| US11793823B2 (en) | 2020-04-23 | 2023-10-24 | Syneurx International (Taiwan) Corp. | Compounds and pharmaceutical uses thereof |
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