WO2014084743A1

WO2014084743A1 - Dendritic core compounds

Info

Publication number: WO2014084743A1
Application number: PCT/NZ2013/000215
Authority: WO
Inventors: Peter Charles Tyler; Olga Vladimirovna Zubkova
Original assignee: Callaghan Innovation Research Ltd
Current assignee: Callaghan Innovation Research Ltd
Priority date: 2012-11-28
Filing date: 2013-11-28
Publication date: 2014-06-05
Anticipated expiration: 2015-05-28
Also published as: CN104981455B; EP2925722A1; EP2925722A4; JP2016506382A; CN104981455A; US20150291522A1; US9828341B2; JP6445453B2

Abstract

The invention relates to compounds that are useful for the preparation of dendrimer compounds, the use of these compounds for preparing dendrimers and processes for preparing the compounds.

Description

DENDRITIC CORE COMPOUNDS

TECHNICAL FIELD This invention relates generally to compounds that are intermediates for the preparation of dendrimer compounds, the use of these compounds for preparing dendrimers and processes for preparing the compounds.

BACKGROUND

Interest in dendritic compounds has grown in recent times, and dendrimers have found application in a wide range of areas, such as pharmaceuticals, drug delivery, gene delivery, sensor technologies (cation photodetection and fluorescence signal quenching), in the synthesis of monodisperse metallic nanoparticles, in environmental remediation, as blood substitutes usually in the oxygen-carrying sense (oxygen therapeutics) and plasma substitutes (volume expenders).

Dendritic structures tend to be complex. However, for some applications, such as pharmaceuticals, it is desirable to employ compounds that are discrete, well-characterised, single entities. This can be an issue in the dendrimer space, so it is desirable to use intermediates and processes which produce such single entities. There is therefore a need for simple dendritic cores that can be used to prepare dendritic compounds for use in certain applications. Newkome et al. (Macromolecules, 1993, 26, 2394-2396) have described the synthesis of a dodecaacid dendritic core, starting from the tetraacid compound (structure shown in Scheme 1 , below) and coupling this with a branched amine. This produced a compound with twelve terminal acids (attachment points), which was further elaborated (through five generations) to produce dendrimers having up to 972 terminal acids.

Newkome et al. {J. Org. Chem. 2002, 67, 3957-3960) have also reported the synthesis of a tetraamine (amine terminated) dendritic core, starting from the tetraacid compound. The tetraamine core was further elaborated by reaction with excess acrylonitrile to generate an octanitrile as an oily product.

Hukkamaki et al. (Hukkamaki, J.; Pakkanen, T.T. Journal of Molecular Catalysis A: Chemical 2001 , 174, 205-21.) have also described the synthesis of the same amine terminated dendritic core. The tetraamine core was further elaborated by reaction with acrylonitrile to produce a (two generation) dendrimer having eight terminal amine groups.

For the synthesis of simple, well-defined dendritic molecules as single compounds, it would be desirable to employ a simple, stable dendritic core as an intermediate which could provide a route to the synthesis of, for example, dendrimers having pharmaceutical uses.

It is therefore an object of the present invention to provide intermediate compounds for the preparation of dendritic compounds, or to at least provide a useful choice.

SUMMARY OF INVENTION

In a first aspect, the invention provides a compound of formula (I)

wherein:

Y is O;

B is O;

R¹ and R² are absent; and

either A, E, D and X are all CH₂; or A, D and X are all CH₂ and E is (CH₂CH₂0)t CH2 wherein indicates a point of attachment of E to its adjacent carbonyl group;

t is an integer from 1 to 10; or wherein:

Y is C;

R¹ and R² are both H; and

A, E, B and D are CH₂ and X is O; or wherein:

Y is C;

R and R² are both H;

B, X, D and E are all absent; and u is an integer from 1 to 10; or wherein:

Y is C;

X is O;

B is (CH₂)_P;

A, E and D are all CH₂; and

R is H, NHZ or Ci_-6alkyl and R² is a radical of formula (i) or a radical of formula (ii)

Z is H, acyl, C(0)(CH₂)wN(H)G, ^*CH₃ ^*C(0)- where ^*C denotes ³C or ¹⁴C, 5-TAMRA (4- carboxytetramethylrhodamine), Fluorescein (resorcinolphthalein), Alexa Fluor 350 (7-amino-4- methyl-6-sulfocoumarin-3-acetic acid), BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) or Alkyne MegaStokes dye 608 (1-{3-{[4-(2-cyclooctyn-1-ylmethyl)benzoyl]amino}propyl}-4-{2-[4- (dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate);

w is an integer from 1 to 11 ;

G is H, acyl, Boc (t-butoxycarbonyl), Troc (2,2,2-trichloroethyloxycarbonyl), Fmoc (9- fluorenylmethoxycarbonyl), Cbz (benzyloxycarbonyl),^*CH₃ ^*CO- where ^*C denotes ¹³C or ¹⁴C, 5- TAMRA (4-carboxytetramethylrhodamine), Fluorescein (resorcinolphthalein), Alexa Fluor 350 (7-amino-4-methyl-6-sulfocoumarin-3-acetic acid), BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s- indacene) or Alkyne MegaStokes dye 608 (1-{3-{[4-(2-cyclooctyn-1- ylmethyl)benzoyl]amino}propyl}-4-{2-[4-(dimethylamino)phenyl]ethenyl}pyridinium

hexafluorophosphate); or wherein:

Y is C;

X is O;

B is (CH₂)_P;

A, E and D are all CH₂; and

R and R², both the same, are a radical of formula (i) or a radical of formula (ii)

R³ is a radical of formula (iii) or a radical of formula (iv)

(iii) (iv) each T is independently selected from the group consisting of (CH₂CH₂0)_XCH₂CH₂ and CH₂; each x is independently an integer from 1 to 12; m is an integer from 1 to 11 , provided that when T is (CH₂CH₂0)xCH2CH₂ then m is 1 ; n is an integer from 1 to 11 , provided that when T is (CH₂CH₂0)xCH₂CH₂ then n is 1 ; p is an integer from 1 to 5;

R⁵ is H,

; and M is sodium or ammonium.

Preferably each T is CH₂.

Alternatively preferably at least one T is CH₂.

Alternatively preferably at least one T is (CH₂CH₂0)xCH₂CH₂.

More preferably each T is CH₂ and m is an integer from 3 to 10, e.g. an integer from 4 to 9, e.g. an integer from 5 to 8, e.g. an integer from 6 to 7. Most preferably m is 7. Preferably each T is CH₂ and n is an integer from 3 to 10, e.g. an integer from 4 to 9, e.g. an integer from 5 to 8, e.g. an integer from 6 to 7. Most preferably n is 7.

Alternatively preferably at least one T is CH₂ and m is an integer from 3 to 10, e.g. an integer from 4 to 9, e.g. an integer from 5 to 8, e.g. an integer from 6 to 7. Most preferably m is 7. Preferably at least one T is CH₂ and n is an integer from 3 to 10, e.g. an integer from 4 to 9, e.g. an integer from 5 to 8, e.g. an integer from 6 to 7. Most preferably n is 7. Alternatively preferably at least one T is (CH2CH₂0)xCH₂CH₂ and x is an integer from 2 to 10, e.g. an integer from 2 to 9, e.g. an integer from 2 to 8, e.g. an integer from 2 to 7, e.g. an integer from 2 to 6, e.g. an integer from 2 to 5, e.g. an integer from 2 to 4. Most preferably x is 3. Preferably Y is O. Alternatively, it is preferred that Y is C.

R⁵-0 ^'.V o ^TV

Preferably R³ is a radical of formula (iii) (iii)

Alternatively it is preferred that R³ is a radical of formula (iv) (^iv)

It is further preferred that, in R³, one T is (CH₂CH₂0)xCH₂CH₂ and one T is CH₂.

It is further preferred that, in R³, (T)_m is (CH₂CH20)xCH2CH₂ and (T)_n is (CH₂)_n. Alternatively it is preferred that, in R³, (T)„ is (CH₂CH20)xCH₂CH2 and (T)_m is (CH₂)_m.

Alternatively preferably R³ is a radical of formula (iv) wherein each T is (CH₂CH₂0)xCH₂CH2, wherein each x in each radical of formula (iv) is independently selected.

Preferably R¹ and R² are both a radical of formula (i) (i)

Alternatively it is preferred that R¹ and R² are both a radical of formula (ii)

It is further preferred that, in R¹, one T is (CH₂CH₂0)xCH2CH₂ and one T is CH₂ and, in R², one T is (CH₂CH20)_XCH₂CH2 and one T is CH₂. It is further preferred that, in both R¹ and R², (T)_m is (CH₂CH₂0)_XCH₂CH₂ and (T)„ is (CH₂)_n. Alternatively it is preferred that, in both R¹ and R², (T)_n is (CH₂CH20)xCH2CH2 and (T)_m is (CH₂)_m. Alternatively it is preferred that, in both R¹ and R², each T is (CH₂CH₂0)_xCH₂CH₂, wherein each x in each radical of formula (ii) is independently selected.

Alternatively it is preferred that R is H or Ci_-6alkyl, e.g. CH₃ or CH₂CH₃. Alternatively it is preferred that R¹ is NH₂. Alternatively it is preferred that R is NHZ, more preferably where Z is C(0)(CH₂)_wN(H)G, e.g. where G is Troc (2,2,2-trichloroethyloxycarbonyl), Fmoc (fluorenylmethyloxycarbonyl) or Cbz (benzyloxycarbonyl). Preferably w is 7.

Preferably R⁵ is selected from the group consisting of

Alternatively preferably R is H. Alternatively preferably R is

Preferably Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, and X are all CH₂ and E is (CH₂CH₂0)_tCH₂; and t is an integer from 1 to 10, preferably integer from 1 to 2.

Alternatively it is preferred that Y is C; R¹ and R² are both H; A, E, B and D are CH₂ and X is O.

Alternatively it is preferred that Y is C; A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; and u is an integer from 1 to 10.

Alternatively it is preferred that:

Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P;

R¹ is H, NHZ or d_-6alkyl;

R² is a radical of formula (i) or a radical of formula (ii) (i)

Z is H, acyl, C(0)(CH₂)wN(l-l)G, ^*CH₃ ^*CO- where ^*C denotes ¹³C or ¹⁴C, 5-TAMRA (4- carboxytetramethylrhodamine), Fluorescein (resorcinolphthalein), Alexa Fluor 350 (7-amino-4- methyl-6-sulfocoumarin-3-acetic acid), BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) or Alkyne MegaStokes dye 608 (1-{3-{[4-(2-cyclooctyn-1-ylmethyl)benzoyl]amino}propyl}-4-{2-[4- (dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate);

w is an integer from 1 to 1 1 ;

G is H, acyl, (t-butoxycarbonyl), Troc (2,2,2-trichloroethyloxycarbonyl), Fmoc (9- fluorenylmethoxycarbonyl), Cbz (carboxybenzyl),^{* *}CH₃ ^*CO- where ^*C denotes ¹³C or ¹⁴C, 5- TAMRA (4-carboxytetramethylrhodamine), Fluorescein (resorcinolphthalein), Alexa Fluor 350 (7-amino-4-methyl-6-sulfocoumarin-3-acetic acid), BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s- indacene) or Alkyne MegaStokes dye 608 (1-{3-{[4-(2-cyclooctyn-1- ylmethyl)benzoyl]amino}propyl}-4-{2-[4-(dimethylamino)phenyl]ethenyl}pyridinium

hexafluorophosphate).

Preferably R² is a radical of formula (ii) and R is H, NHZ or Ci_-6alkyl. It is further preferred that, in R², one T is (CH₂CH₂0)_XCH2CH₂ and one T is CH₂. It is further preferred that, in R², (T)_m is (CH₂CH₂0)_xCH₂CH₂ and (T)„ is (CH₂)_n. Alternatively it is preferred that, in R², (T)_n is (CH₂CH₂0)_xCH₂CH₂ and (T)_m is (CH₂)_m. Alternatively it is preferred that, in R², each T is (CH₂CH₂0)_xCH₂CH₂, wherein each x in each radical of formula (ii) is independently selected. Alternatively it is preferred that Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P; R¹ and R², both the same, are a radical of formula (i) or a radical of formula (ii)

(i) (ii)

Preferably, Y is C; X is is (CH₂)_P; R¹ and R², both the same, are a

radical of formula (i) ; R³ is a radical of formula (iii)

; and p is 1.

Alternatively preferably Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P; R¹ and R², both the

same, are a radical of formula (ii) (") ; R³ is a radical of

formula (iv) (^iv) ; and p is 1. Preferably Y is C; R¹ and R² are both H; A, E, B and D are CH₂; X is O; and R³ is a radical of

^RS"W o

formula (iii) ('»>

Alternatively preferably Y i and R² are both H; A, E, B and D are CH₂; X is O; and R³ is a

radical of formula (iv)

Preferably Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P; R¹ is H; R² is a radical of formula (i) ; R³ is a radical of formula (iii)

('") ; and p is 1.

Alternatively pref all CH₂; B is (CH₂)_P; R¹ is H; R² is a radical

of formula (ii) (ii) R³ is a radical of formula (iv)

(iv) ; and p is 1.

Alternatively preferably Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P; R¹ is H; R² is a radical

of formula (ii) ; R³ is a radical of formula (iii)

and p is 1.

Alternatively pref D are all CH₂; B is (CH₂)_P; R is H; R² is a radical

of formula (i) (i) and R³ is a radical of formula

(iv) ; and p is 1.

Preferably Y is C; A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; u is an integer

from 1 to 10; and R³ is a radical of formula (iii) (iii) Alternatively preferably Y is C; A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; u

is an integer from 1 to 10; and R³ is a radical of formula (iv) (iv) Preferably Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)_tCH₂; t is an integer from 1 to 10, preferably an integer

from 1 to 2; and R³ is a radical of formula (iii)

Alternatively preferably Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)_tCH₂; t is an integer from 1 to 2; and R³ is a

radical of formula (iv) ^(IV) Preferably p is 1 . Preferably t is an integer from 1 to 2. Preferably M is sodium.

Preferably the compound of formula (I) is selected from the group consisting of:

11



5





In another aspect, the invention provides a compound selected from the group consisting of:

In another aspect, the invention provides the use of a compound of formula (I) as defined above for preparing a dendritic compound.

In still another aspect, the invention provides a method of preparing a dendritic compound, including the step of contacting a compound of formula (I) with a glycoside compound that contains a free amino group.

It will be appreciated that any of the sub-scopes disclosed herein, e.g. with respect to A, B, D, E, G, X, R¹, R², R³, R⁴, R⁵, n, m, p t, u, w, x, T, Y and Z may be combined with any of the other sub-scopes disclosed herein to produce further sub-scopes. DETAILED DESCRIPTION

Definitions The term "CrC₆alkyl" means any saturated hydrocarbon radical having up to 6 carbon atoms and is intended to include both straight- and branched-chain alkyl groups. Examples of alkyl groups include: methyl group, ethyl group, n-propyl group, /^'so-propyl group, n-butyl group, iso- butyl group, sec-butyl group, f-butyl group, n-pentyl group, 1 ,1-dimethylpropyl group, 1 ,2- dimethylpropyl group, 2,2-dimethylpropyl group, 1-ethylpropyl group, 2-ethylpropyl group, n- hexyl group and 1-methyl-2-ethylpropyl group.

The term "acyl" means C(=0)R' group, where R' is a CrC₃₀alkyl group, where Ci-C₃₀alkyl means any saturated hydrocarbon radical having up to 30 carbon atoms, and is intended to include straight chain alkyl groups. Examples include acetyl group.

Compounds of the Invention

The compounds of the invention are simple, easy to use "star burst PET-PEG" dendritic cores. They are useful for the preparation of dendrimers, thereby providing a facile route to well- defined cluster molecules, e.g. pharmaceutical cluster molecules, as single compounds. The compounds of the invention are two-, three- or four-directional short-armed and long-armed dendritic cores. Some compounds of the invention are "pre-activated" (e.g. those where R⁵ is a succinimidyl group) so that they can be used, without coupling reagents, for synthesising dendritic compounds. The dendritic core "arms" can be elongated in a controlled manner with suitable linkers, such as 6-8-carbon linkers or PEG linkers (one or two generations), making the "arms" shorter or longer as desired. The linkers are flexible and the tetrahedral model of the core is stable. Compounds of the invention that are succinimidyl esters are, advantageously, crystalline compounds. They are therefore stable and can be easily stored. Conveniently, the compounds of the invention can be purified by flash chromatography. Because of their simplicity of design, the compounds provide a novel approach to reaching larger sized dendrimers, but employing fewer "arms" (2-4 "arms" instead of 32-64), thus simplifying the dendrimer product. Dendritic cores with long carbon or PEG chains can be considered to be mimics of carbohydrate chains, but they avoid the costs and synthetic challenges associated with preparing linear oligosaccharides. The invention relates to:

1. A compound of formula (I)

wherein:

Y is O;

B is O;

R¹ and R² are absent; and

either A, E, D and X are all CH₂; or A, D and X are all CH₂ and E is (CH₂CH₂0)t^#CH2 wherein * indicates a point of attachment of E to its adjacent carbonyl group;

t is an integer from 1 to 10; or wherein:

Y is C;

R¹ and R² are both H; and

A, E, B and D are CH₂ and X is O; or wherein:

Y is C;

A is (CH₂)_U

R¹ and R² are both H;

B, X, D and E are all absent; and

u is an integer from 1 to 10; or wherein:

Y is C;

X is O;

B is (CH₂)_P;

A, E and D are all CH₂; and

R¹ is H, NHZ or Ci_-6alkyl and R² is a radical of formula (i) or a radical of formula (ii)

Z is H, acyl, C(0)(CH₂)_wN(H)G₎ ^*CH₃ ^*C(0)- where ^*C denotes ¹³C or ¹⁴C, 4- carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6-sulfocoumarin-3- acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3-{[4-(2-cyclooctyn-1- ylmethyl)benzoyl]amino}propyl}-4-{2-[4-(dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate; w is an integer from 1 to 11 ;

G is H, acyl, t-butoxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 9- fluorenylmethoxycarbonyl, benzyloxycarbonyl,^*CH₃ ^*CO- where ^*C denotes ¹³C or ¹ C, 4- carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6-sulfocoumarin-3- acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3-{[4-(2-cyclooctyn-1- ylmethyl)benzoyl]amino}propyl}-4-{2-[4-(dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate; or wherein:

Y is C;

X is O;

B is (CH₂)_P;

A, E and D are all CH₂; and

R¹ and R², both the same, are a radical of formula (i) or a radical of formula (ii)

R³ is a radical of formula (iii) or a radical of formula (iv)

(iii) (iv) each T is independently selected from the group consisting of (CH₂CH₂0)_XCH₂CH₂ and CH₂; each x is independently an integer from 1 to 12; m is an integer from 1 to 11 , provided that when T is (CH₂CH20)_XCH₂CH₂ then m is 1 ; n is an integer from 1 to 11 , provided that when T is (CH₂CH₂0)xCH₂CH₂ then n is 1 ; p is an integer from 1 to 5;

M is sodium or ammonium.

2. A compound as described in the above paragraph 1 wherein each T is CH₂. 3. A compound as described in the above paragraph 1 wherein at least one T is

4. A compound as described in the above paragraph 1 wherein m is an integer from 5 to 8. 5. A compound as described in the above paragraph 1 or 2 wherein n is an integer from 5 to

6. A compound as described in any one of the above paragraphs 1 to 5 wherein Y is C. 7. A compound as described in any one of the above paragraphs 1 to 6 wherein R³ is a radical of formula (iii)

8. A compound as described in any one of the above paragraphs 1 to 6 wherein R³ is a radical of formula (iv)

(iv)

A compound as described in any one of the above paragraphs 1 to 8 wherein R¹ and R² are both a radical of formula (i)

A compound as described in any one of the above paragraphs 1 to 8 wherein R¹ and R² are both a radical of formula (ii)

A compound as described in any one of the above paragraphs 1 to 8 wherein R is H Ci_-6alkyl.

A compound as described in any one of the above paragraphs 1 to 8 wherein R¹ is NH₂. pound as described in any one of the above paragraphs 1 to 12 wherein R⁵

A compound as described in any one of the above paragraphs 1 to 5, 7 to 8 or 13 wherein Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)_tCH₂; and t is an integer from 1 to 10.

A compound as described in any one of the above paragraphs 1 to 8 or 13 wherein Y is C; R¹ and R² are both H; A, E, B and D are CH₂ and X is O.

A compound as described in any one of the above paragraphs 1 to 8 or 13 wherein Y is C; A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; and u is an integer from 1 to 10. A compound as described in any one of the above paragraphs 1 to 8 or 13 wherein Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P;

R¹ is H, NHZ or C_1-6alkyl;

R² is a radical of formula i) or a radical of formula (ii)

0) (ii)

Z is H, acyl, C(OXCH₂)wNH(G), *CH₃*CO- where *C denotes ¹³C or ¹⁴C, 4- carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6- sulfocoumarin-3-acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3- {[4-(2-cyclooctyn-1-ylmethyl)benzoyl]amino}propyl}-4-{2-[4- (dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate;

w is an integer from 1 to 1 1 ;

G is H, acyl, t-butoxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 9- fluorenylmethoxycarbonyl, benzyloxycarbonyl, *CH₃*CO- where *C denotes ¹³C or ¹⁴C, 4-carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6- sulfocoumarin-3-acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3- {[4-(2-cyclooctyn-1-ylmethyl)benzoyl]amino}propyl}-4-{2-[4- (dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate. A compound as described in any one of the above paragraphs 1 to 9 or 13 wherein, Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P; R¹ and R², both the same, are a radical of formula (i)

(i)

R³ is a radical of formula (iii)

^RS-W o

; and and p is 1. A compound as described in any one of the above paragraphs 1 to 8 or 10 or 13 wherein

Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P; R¹ and R², both the same, are a radical of formul

and R³ i

(iv)

and p is 1. A compound as described in any one of the above paragraphs 1 to 8 or 13 or 15 wherein

Y is C; R¹ and R² are both H; A, E, B and D are CH₂; X is O; and R³ is a radical of formula (iii)

^R5"° o ^TV

(iii) A compound as described in any one of the above paragraphs 1 to 8 or 13 or 15 wherein

Y is C; R¹ and R² are both H; A, E, B and D are CH₂; X is O; and R³ is a radical of formula (iv)

(iv)

A compound as described in any one of the above paragraphs 1 to 8 or 13 or 17 wherein

Y is C; X B is (CH₂)_P; R¹ is H; R² is a radical of formula (i)

(i)

R³ is a radical of formula (iii)

(iii)

and p is 1. A compound as described in any one of the above paragraphs 1 to 8 or 13 or 17 wherein Y is C; X (CH₂)_P; R¹ is H; R² is a radical of formula (ii)

(ii)

R³ is a radical of formula (iv)

and p is 1.

Y is C; X (CH₂)_P; R¹ is H; R² is a radical of formula (ii)

(ii)

R³ is a radical of formula (Hi)

(iii)

and p is 1.

Y is C; X B is (CH₂)_P; R is H; R² is a radical of formula (i)

(i)

and R³ is a radical of formula (iv)

and p is 1. A compound as described in any one of the above paragraphs 1 to 8 or 13 or 16 wherein

Y is C; A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; u is an integer from 1 to 10; a ³ is a radical of formula (iii)

(iii) A compound as described in any one of the above paragraphs 1 to 8 or 13 or 16 wherein

Y is C; A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; u is an integer from 1 to 10; a ³ is a radical of formula (iv)

(iv) A compound as described in any one of the above paragraphs 1 to 8 or 13 or 14 wherein

Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)tCH2; t is an integer from 1 to 2; and R³ is a radical of formula (iii)

(iii) A compound as described in any one of the above paragraphs 1 to 8 or 13 or 14 wherein

Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)_tCH₂; t is an integer from 1 to 2; and R³ is a radical of formula (iv

(iv) A compound as described in any one of the above paragraphs 1 to 10, 17 to 19 or 22 to 25 wherein p is . A compound as described in any one of the above paragraphs 1 to 30 wherein M is sodium. A compound as described in the above paragraph 1 , selected from the group consisting of:

30

32

33. A compound selected from the group consisting of:

Synthesis of the Compounds of the Invention

The compounds of the invention may be prepared by a variety of different methods. The following are representative non-limiting general methods for synthesising compounds of the invention.

Those skilled in the art will realise that Schemes 2 to 7 below show compounds of the invention where R⁵ is succinimidyl group. However, compounds of the invention where R⁵ is other than succinimidyl can be prepared analogously. For example, compounds where R⁵ is H can be prepared by treatment of the OBn precursors (e.g. Schemes 2-7, below) with palladium on carbon or palladium hydroxide on carbon or platinum on carbon catalysts in solvents such as aqueous THF, methanol, ethanol, ethyl acetate, stirred under a hydrogen atmosphere at ambient temperature and pressure or at 5-50 psi, preferably at 5-25 psi. "Tetrameric" dendritic core compounds of the invention (where R¹ and R² are both a radical of formula (i) or a radical of formula (ii)) are prepared from compound 1 , which is synthesised in three steps from pentaerythritol (Scheme 1 ).

Scheme 1

(Ref.1 , Hukkamaki, J.; Pakkanen, T.T. Journal of Molecular Catalysis A: Chemical 2001 , 174, 205-21 1 ; Ref. 2, Newcombe, G. R.; Mishra, A; Moorfield, C. N. J. Org. Chem. 2002, 67, 3957- 3960).

Compound 1 is then converted to compounds of formula (I) via reaction with a suitable amino- substituted carboxylic acid, such as 3-aminopropanoic acid, 4-aminobutanoic acid, 5- aminopentanoic acid, 6-aminohexanoic acid, 7-aminoheptanoic acid, 8-aminooctanoic acid, 9- aminononanoic acid, 10-aminodecanoic acid, 11 -aminoundecanoic acid or 12-aminododecanoic acid (Scheme 2). cheme 2

"Dimeric" dendritic core compounds of the invention where Y is C and R¹ and R² are both H are prepared from 3,3'-(propane-1 ,3-diylbis(oxy)dipropanoic acid, using a suitable amino- substituted carboxylic acid, such as 3-aminopropanoic acid, 4-aminobutanoic acid, 5- aminopentanoic acid, 6-aminohexanoic acid, 7-aminoheptanoic acid, 8-aminooctanoic acid, 9- aminononanoic acid, 10-aminodecanoic acid, 11-aminoundecanoic acid or 12-aminododecanoic acid (Scheme 3). Scheme 3

'-(propane-1 ,3-diylbis(oxy))dipropanoic acid

H₂N jj^ BnOH o

SOCI, "Π OBn

n is an integer from 1 to 11

Η,Ν BnOH

^H _rSc ^H2N"(T Bn

'2 )rf ^O

m is an integer from 1 to 11

1. Pd/C

2. EDC, NHS

"Dimeric" dendritic core compounds of the invention where Y is C; A is (CH₂)_U; and R¹ and R² are both H are prepared from a suitable diacid, as shown in Scheme 4. Scheme 4

Diacid

H₂N_ _ II

where T = (CH₂CH₂0)_x-CH₂CH₂ (CH₂) θ^η' ^OBn

Et₃N

an integer from 1 to 11

^H2 )m^OBn

m is an integer from 1 to 11

"Dimeric" dendritic core compounds of the invention where Y is O and E is (CH2CH₂0)_tCH2 are prepared as shown in Scheme 5. Suitable starting materials include, for example, 3,6,9- trioxaundecanedioic acid, 3,6,9,12-tetraoxatetradecanedioic acid or 3,6,9,12,15- pentaoxaheptadecanedioic acid. Scheme 5

"Trimeric" dendritic core compounds of the invention where Y is C and R¹ is NHZ are prepared from 2-amino-2-hydroxymethyl-propane-1 ,3-diol, as shown in Scheme 6. Those skilled in the art will appreciate that trimeric dendritic core compounds where R¹ is NH₂ can be used to further elaborate the substitution at the R¹ position. For example, such compounds can be linked to radio- or fluorescent labels or the amino functionality can be converted to other functional groups.

Scheme 6

"Trimeric" dendritic core compounds of the invention where Y is C and R¹ is H, CH₃ or CH₂CH₃ are prepared from 2-hydroxymethyl-propane-1 ,3-diol, 1 ,1 ,1-tris(hydroxymethyl)ethane or 1 ,1 ,1- tris(hydroxymethyl)propane, respectively (Scheme 7).

Scheme 7

where T = (CH₂CH₂0)_x-CH₂CH₂ or _(CH₂)

1. NH₂(CH₂)_mC0₂Bn

2. Pd(OH)₂/C

n is an integer from 1 to 11

3. EDC, NHS

m is an integer from 1 to 11

Use of the Compounds of the Invention for Preparing Dendrimers

The compounds of the invention are useful for the preparation of dendritic compounds. Conveniently, the compounds of the invention allow for facile synthesis of dendrimers. Advantageously, with some compounds of the invention there is no need to use coupling reagents and then to remove an excess of reagents from the dendritic products. The coupling procedure is simple and requires a suitable solvent (DMF, DMSO, water, for example), a small amount of base, e.g. triethylamine, and any glycoside with a free amino group (at least about 2 equivalents of glycoside, e.g. about 2.2 equivalents of glycoside are used for coupling with dimeric compounds of the invention, at least about 3 equivalents of glycoside, e.g. about 3.3 equivalents of glycoside are used for coupling with trimeric compounds of the invention and at least about 4 equivalents of glycoside e.g. about 4.4 equivalents of glycoside are used for coupling with tetrameric compounds of the invention).

Using this method, it is possible to attach a variety of carbohydrate fragments (e.g. heparan sulfates, sulfated monosaccharides, oligosaccharides, amino acids, radioligands, imaging agents, fluorescent probes, antibiotics, cytostatic drugs (chemotherapy), veterinary pharmaceuticals, proton and pH sensors, metal ions and ferrocene) with various linkers to the dendritic cores. By way of example, Scheme 8 shows how a "star burst PET-PEG" dendritic cluster glycomimetic of heparan sulfate can be synthesised from a dendritic core of the present invention. This heparan sulfate mimetic is useful as an inhibitor of Alzheimer's β-secretase.

cheme 8

Those skilled in the art will appreciated that, using the tetrameric compounds of the invention, it is possible to achieve dendrimers that are a similar size to 16-mer and 32-mer ball-clusters, but which have fewer copies of glycosides attached. Typically, when the use of 16-, 32- and 64- mer dendritic cores is reported, the dendrimer products are described as mixtures with only partial capping of attachment points due to electrostatic repulsion and steric crowding effects. Using the compounds of the invention, with fewer linkers, the glycosides are well separated in space so the capping leads to fully substituted dendrimers. This is advantageous, for example, in the field of pharmaceuticals where it is desirable to produce discrete, well- characterised chemical compounds, rather than mixtures. ABBREVIATIONS

NMR Nuclear magnetic resonance

HRMS High resolution mass spectrometry

DCM Dichloro methane

DMF N,N-Dimethylformamide

EtOH Ethanol

eOH Methanol

THF Tetrahydrofuran

EtOAc Ethyl acetate

RT Room temperature

TAMRA 4-Carboxytetramethylrhodamine N-succinimidyl ester

BODIPY 4,4-Difluoro-4-bora-3a,4a-diaza-s-indacene

EDC 1 -Ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride

NHS N-Hydroxysuccinimide

EXAMPLES

The following examples further illustrate the invention. It is to be appreciated that the invention is not limited to the examples.

Example 1: Synthesis of Compounds of the Invention

cheme 9

Preparation of 1

Tetranitrile precursor (Ref.1 , Hukkamaki, J.; Pakkanen, T.T. Journal of Molecular Catalysis A: Chemical 2001 , 174, 205-211 ) is prepared via Michael-type addition of acrylonitrile to pentaerythritol. Acidic hydrolysis of tetranitrile (Ref. 2, Newcombe, G. R.; Mishra, A; Moorfield, C. N. J. Org. Chem. 2002, 67, 3957-3960) furnishes the tetraacid. Tetraacid (1.0 g, 2.35 mmol) is dissolved in dry DMF (15 mL). N-Hydroxysuccinimide (1 .62 g, 14.14 mmol) and 1 -ethyl-3-[3- dimethylaminopropyl]carbodiimide hydrochloride (EDC, 2.71 g, 14.14 mmol) are added to the reaction mixture at room temperature and stirring continued for 24 hrs. The mixture is diluted with DCM and washed with water, then with diluted HCI and water, dried over magnesium sulfate and concentrated. The residue is purified by flash chromatography on silica gel eluting with EtOAc followed by EtOAc:MeOH, 19:1→ 9:1 → 7:1→ 4:1 to give the tetra-succinimidyl ester (1 , 1.2 g, 1.48 mmol, 63%). R_f = 0.25 (Ethyl Acetate :MeOH, 9:1 ). ¹³C-N R (125MHz, DMSO-D₆) δ 170.7, 170.1 , 68.7, 65.5, 44.9, 31.5, 25.7. HRMS calcd for C₃3H4oN₄0₂oNa (M+Naf m/z 835.2134, found 835.2128.

Preparation of 2

Aminooctanoic acid is treated with benzyl alcohol in the presence of thionyl chloride (Ref. 3, Patel, R. P; Price, S. J. Org Chem. 1965, 30 (10), 3575-3576) to give a tetra benzyl ester (2 g, 8.12 mmol). This and tetra-succinimidyl ester (1, 1.1 g, 1.35 mmol) are dissolved in a mixture of dry THF (55 mL) and dry DMF (3 mL) and treated with triethylamine (1.5 mL, 10.83 mmol). After stirring for 24 hrs the mixture is diluted with ethyl acetate and washed with water twice, dried over magnesium sulfate and concentrated. The residue is dissolved in hot EtOAc, the crystals are filtered off and discharged. The mother liquor is concentrated and the residue is purified by flash chromatography on silica gel eluting with Chloroform : EtOAc : MeOH, 5:2:0.5 to afford the tetra-benzyl ester (2, 1.7 g, 1.26 mmol, 93%). R_f = 0.3 (Chloroform : Ethyl Acetate : MeOH, 5:2:1 ). ¹³C-NMR (125MHz, CDCI₃) δ 173.5, 171.2, 136.1 , 128.5, 128.2, 128.1 , 69.1 , 67.4, 66.1 , 45.3, 39.5, 36.9, 34.2, 29.6, 29.4, 28.9, 26.7, 26.6, 24.8. HRMS calcd for C₇₇Hii₂N₄0i₆Na (M+Naf m/z 1371.7971 , found 1371.7977. Preparation of 3

Tetra benzyl ester (2, 0.595 g, 441 μηιοΙ) is dissolved in dry THF (16 mL). Water (4 mL) and glacial acetic acid (5 drops) are added. The reaction mixture is treated with palladium hydroxide on carbon (20% Pd, 1 g) and stirred for 3 hours under hydrogen at ambient temperature and pressure. The catalyst is filtered off and washed with 50% aqueous EtOH. The solution is concentrated to dryness to give a "long-armed" tetraacid (3, 0.42 g, 429 pmol, 97%). The product is used in the next step without further purification. R_f = 0.0 (base line, Chloroform : Ethyl Acetate : MeOH, 5:2:1 ). ¹³C-NMR (125MHz, DMSO-D₆) δ 174.5, 169.9, 68.8, 67.3, 45.0, 39.0, 38.4, 36.1 , 33.8, 29.1 , 28.5, 38.4, 26.3, 25.2, 24.5. HRMS calcd for C₄₉H₈₇N₄Oi₆ (M-HV m/z 987.6117, found 987.6110.

Preparation of 4

"Long-armed" tetraacid (3, 424 mg, 429 mol) is dissolved in dry DMF (7 mL). N-

Hydroxysuccinimide (296 mg, 2.57 mmol) and 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC, 493 mg, 2.57 mmol) are added to the reaction mixture at room temperature and stirring continued for 24 hrs. The mixture is diluted with DCM and washed with water, then with diluted HCI and water, dried over magnesium sulfate and concentrated. The residue is purified by flash chromatography on silica gel eluting with Chloroform : Ethyl Acetate : MeOH, 5:2:0.5→ 5:4: 1 to give the "long-armed" tetra-succinimidyl ester (4, 415 mg, 301 μιηοΙ, 70.3 %). R_f = 0.25 (DCM:MeOH, 9: 1 ). ³C-NMR (125MHz, CDCI₃) δ 171.5, 169.4, 168.6, 68.9, 67.4, 45.3, 39.4, 36.7, 30.8, 29.5, 29.4, 28.6, 28.5, 26.6, 25.5, 25.4, 24.4. HRMS calcd for

(M+Na)⁺ m/z 1399.6748, found 1399.6737.

Preparation of 5

H₂N(PEG)₃CH₂CH₂COOH (or PEG aminoacid) (1 .0 g, 4.52 mmol) is dissolved in benzyl alcohol (30 mL, 287 mmol) and cooled to 0°C. Thionyl chloride (6 mL, 82.2 mmol) is added slowly dropwise. The reaction mixture is stirred at 0°C for 15 min followed by heating at 100°C for 5 hours. Then this is diluted with diethyl ether and the oily residue is collected and purified by flash chromatography on silica gel eluting with Dichloro methane: MeOH, 9:1 → 1 :1 to afford the benzyl ester (5, 1 .2 g, 3.9 mmol, 85%). R_f = 0.15 (Dichloromethane: MeOH, 9:1 ). ¹³C-NMR (125MHz, CDCI₃) 6 171 .6, 135.8, 128.5, 128.2, 128.1 , 70.2, 70.14, 70.13, 69.9, 66.7, 66.4, 66.3, 50.0, 39.7, 35.0. HRMS calcd for Ci₆H₂6N0₅ (M+H)⁺ m/z 312.181 1 , found 312.1806.

Preparation of 6

The benzyl ester 5 (65 mg, 210 μιηοΙ) and tetra-succinimidyl ester (4, 58 mg, 42.1 μηιοΙ) are dissolved in dry DMF (2 mL) and treated with triethylamine (47 pL, 336 pmol). After stirring for 24 hrs the mixture is diluted with ethyl acetate and washed with water twice, dried over magnesium sulfate and concentrated. The residue is dissolved in hot EtOAc, the crystals are filtered off and discharged. The mother liquor is concentrated and the residue is purified by flash chromatography on silica gel eluting with Chloroform : EtOAc : MeOH, 5:2:0.5 to afford the tetra-benzyl ester (6, 71 mg, 32.8 μηιοΙ, 78%). R_f = 0.3 (Chloroform : Ethyl Acetate : MeOH, 5:2: 1 ). ¹³C-NMR (125MHz, CDCI₃) δ 173.2, 171 .3, 135.8, 128.5, 128.2, 128.1 , 70.5, 704, 70.2, 69.9, 69.2, 67.5, 66.5, 66.3, 45.3, 39.4, 39.1 , 36.9, 36.5, 35.1 , 29.6, 29.1 , 29.0, 26.7, 25.5.

Preparation of 7

Tetra benzyl ester (6, 16 mg, 7.21 μιτιοΙ) is dissolved in dry THF (4 mL). Water (1 mL) and glacial acetic acid (2 drops) are added. The reaction mixture is treated with palladium hydroxide on carbon (20% Pd, 20 mg) and stirred for 3 hours under hydrogen at ambient temperature and pressure. The catalyst is filtered off and washed with 50% aqueous EtOH. The solution is concentrated to dryness to give a "long-armed" PEG tetraacid (7, 13 mg, 7.21 pmol, 97%). The product is used in the next step without further purification. R_f = 0.0 (base line, Chloroform : Ethyl Acetate : MeOH, 5:2: 1 ). ¹³C-NMR (125MHz, MeOD) δ 176.7, 174.2, 71 .4, 71 .3, 71 .2, 70.6, 68.8, 67.8, 62.8, 48.5, 40.6, 40.3, 37.8, 37.1 , 35.8, 30.4, 30.2, 30.1 , 30.0, 27.9, 26.9. Preparation of 8

"Long-armed" PEG tetraacid (7, 13 mg, 7.21 pmol) is dissolved in dry DMF (1 mL). N- Hydroxysuccinimide (5.1 mg, 43.2 μιηοΙ), DIPEA (7.6 μί, 43.2 μmol) and -ethyl-3-[3- dimethylaminopropyl]carbodiimide hydrochloride (EDC, 8.3 mg, 43.2 μιηοΙ) are added to the reaction mixture at room temperature and stirring continued for 24 hrs. The mixture is diluted with DCM and washed with water, then with diluted HCI and water, dried over magnesium sulfate and concentrated. The residue is purified by flash chromatography on silica gel eluting with Chloroform : Ethyl Acetate : MeOH, 5:2:0.5→ 5:4:1 to give the "long-armed" PEG tetra- succinimidyl ester (8, 15 mg, 6.85 μιηοΙ, 94 %). R_f = 0.25 (DCM:MeOH, 9:1 ).

Scheme 10

Pd(OH)₂/C/H₂

Preparation of 9

Dodecanedioic acid (1.0 g, 4.34 mmol) is dissolved in dry DMF (10 mL). N-Hydroxysuccinimide (1.51 g, 13.0 mmol, 3 eq.) and 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC, 2.55 g, 13.0 mmol, 3 eq.) and /V,/V-diisopropylethylamine (1.53 mL, 8.68 mmol) are added to the reaction mixture at room temperature and stirring continued for 24 hrs. The mixture is diluted with DCM and washed with water, then with diluted HCI and water, dried over magnesium sulfate and concentrated. The residue is re-crystalized from hot EtOAc to give the di-succinimidyl ester (9, 1.84 g, 4.2 mmol, 98%). ¹³C-NMR (125MHz, DMSO-D₆) δ 170.2, 168.9, 30.2, 28.7, 28.4, 27.9, 25.4, 24.2. HRMS calcd for CzoHzsNsOsNa (M+Na)⁺ m/z 447.1743, found 447.1743. Preparation of 10

Di-succinimidyl ester (9, 500 mg, 1 .18 mmol) and an amino benzyl ester (881 mg, 3.53 mmol, 3 eq.) are dissolved in dry DMF (8 ml.) and treated with triethylamine (0.66 ml_, 4.71 mmol, 4 eq.). After stirring for 24 hrs the mixture is concentrated. The residue is dissolved in hot MeOH, a few drop of chloroform added, the crystals are filtered off and dried to afford the di-benzyl ester (10, 0.75 g, 1 .1 mmol, 92%). ¹³C-NMR (125MHz, MeOD) δ 176.2, 175.5, 137.4, 129.8, 129.4, 129.4, 67.5, 40.6, 37.6, 35.5, 30.7, 30.6, 30.5, 30.2, 30.1 , 27.9, 27.2, 26.1. HRMS calcd for C₄2H64N₂0₆Na (M+Na)⁺ m/z 715.4662, found 715.4664. Scheme 11

HO^O^O^O^OH

Preparation of 14

3,6,9-Trioxaundecanedioic acid (500 mg, 2.25 mmol) is dissolved in dry DMF (10 ml_). N- Hydroxysuccinimide (785 mg, 6.75 mmol, 3 eq.), 1 -ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC, 1 .32 g, 6.75 mmol, 3 eq.) and A/,/V-diisopropylethylamine (2.38 ml_, 13.5 mmol) followed by benzyl 8-aminooctanate (1.68 g, 6.75 mmol, 3 eq.) are added to the reaction mixture at room temperature and stirring continued for 4 hrs. The mixture is diluted with DCM and washed with water, then with diluted HCI and water, dried over magnesium sulfate and concentrated. The residue is dissolved in hot EtOAc, the crystals are filtered off and discharged. The mother liquor is concentrated and the residue is purified by flash chromatography on silica gel eluting with Chloroform : EtOAc : MeOH, 5:2:0.3 to afford the di-benzyl ester (14, 1 .1 g, 1 .61 mmol, 71 %). ¹³C-NMR (125MHz, CDCI₃) δ 173.4, 169.9, 136.1 , 128.4, 128.3, 128.1 , 127.8, 127.7, 65.9, 63.9, 60.3, 39.5, 36.8, 31 .9, 30.9, 29.3, 28.8, 26.6, 25.3, 24.7, 24.4, 21 .2, 20.9. HRMS calcd for CaBHseNzOgNa (M+Na)⁺ m/z 707.3884, found 707.3876. Preparation of 15

Di-benzyl ester (14, 204 mg, 297 pmol) is dissolved in dry THF (8 ml_). Water (2 mL) and glacial acetic acid (3 drops) are added. The reaction mixture is treated with palladium hydroxide on carbon (20% Pd, 0.5 g) and stirred for 3 hours under hydrogen at ambient temperature and pressure. The catalyst is filtered off and washed with 50% aqueous EtOH. The solution is concentrated to dryness to give a "long-armed" diacid (15, 150 mg, 297 μιηοΙ, 99.8%). The product is used in the next step without further purification. R_f = 0.0 (base line, Chloroform : Ethyl Acetate : MeOH, 5:2:1 ). ¹³C-NMR (125MHz, MeOD) δ 178.1 , 175.3, 172.9, 72.3, 71.7, 71.6, 40.4, 35.5, 30.9, 30.6, 30.5, 28.3, 26.5. HRMS calcd for C₂4H4₄N₂0₉Na (M+Na)⁺ m/z 527.2945, found 527.2943.

Preparation of 16

"Long-armed" diacid (15, 150 mg, 297 pmol) is dissolved in dry DMF (4 mL). N- Hydroxysuccinimide (104 mg, 892 μ^ητιοΙ), 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC, 174 mg, 892 μιηοΙ) and Λ/,/V-diisopropylethylamine (0.157 mL, 892 pmol) are added to the reaction mixture at room temperature and stirring continued for 24 hrs. The mixture is diluted with DCM and washed with water, then with diluted HCI and water, dried over magnesium sulfate and concentrated. The residue is purified by flash chromatography on silica gel eluting with Chloroform:Ethyl Acetate : MeOH, 5:2:0.5→ 5:4:1 to give the "long-armed" di- succinimidyl ester (16, 188 mg, 269 pmol, 90 %). R_f = 0.25 (DCM:MeOH, 9:1 ). ¹³C-NMR (125MHz, CDCI₃) δ 170.9, 169.9, 169.5, 168.5, 70.7, 70.6, 70.5, 70.4, 70.0, 38.7, 31.4, 30.7, 29.4, 28.6, 28.5, 28.4, 26.5, 25.5, 24.4. HRMS calcd for C₃2H5oN₄0₁₃ a (M+Naf m/z 721.3272, found 721.3259. Although the invention has been described by way of example, it should be appreciated that variations and modifications may be made without departing from the scope of the invention. Furthermore, where known equivalents exist to specific features, such equivalents are incorporated as if specifically referred to in the specification. INDUSTRIAL APPLICABILITY

Claims

1. A compound of formula (I)

wherein:

Y is O;

B is O;

R¹ and R² are absent; and

either A, E, D and X are all CH₂; or A, D and X are all CH₂ and E is (CH₂CH₂0)t CH2 wherein ^# indicates a point of attachment of E to its adjacent carbonyl group;

t is an integer from 1 to 10; or wherein:

Y is C;

R¹ and R² are both H; and

A, E, B and D are CH₂ and X is O; or wherein:

Y is C;

A is (CH₂)_U

R and R² are both H;

B, X, D and E are all absent; and

u is an integer from 1 to 10; or wherein:

Y is C;

X is O;

B is (CH₂)_P;

A, E and D are all CH₂; and

(■) (ii)

Z is H, acyl, C(OXCH₂)wN(H)G, ^*CH₃ ^*C(0)- where ^*C denotes ¹³C or ¹⁴C, 4- carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6-sulfocoumarin-3- acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3-{[4-(2-cyclooctyn-1- ylmethyl)benzoyl]amino}propyl}-4-{2-[4-(dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate;

w is an integer from 1 to 1 1 ;

G is H, acyl, t-butoxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 9- fluorenylmethoxycarbonyl, benzyloxycarbonyl,*CH₃ ^*CO- where *C denotes ¹³C or ¹⁴C, 4- carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6-sulfocoumarin-3- acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3-{[4-(2-cyclooctyn-1- ylmethyl)benzoyl]amino}propyl}-4-{2-[4-(dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate; or wherein:

Y is C;

X is O;

B is (CH₂)_P;

A, E and D are all CH₂; and

and R², both the same, are a radical of formula (i) or a radical of formula (ii)

(i) (ii) ;

R³ is a radical of formula (iii) or a radical of formula (iv)

(iii) (iv) each T is independently selected from the group consisting of (CH₂CH20)_XCH2CH₂ and CH₂; each x is independently an integer from 1 to 12; m is an integer from 1 to 11 , provided that when T is (CH₂CH₂0)_XCH₂CH₂ then m is 1 ; n is an integer from 1 to 11 , provided that when T is (CH₂CH₂0)_XCH₂CH₂ then n is 1 ; p is an integer from 1 to 5;

M is sodium or ammonium.

2. A compound as claimed in claim 1 wherein each T is CH₂.

3. A compound as claimed in claim 1 wherein at least one T is (CH₂CH₂0)_XCH₂CH₂.

4. A compound as claimed in any one of claims 1 to 3 wherein Y is C.

5. A compound as claimed in any one of claims 1 to 4 wherein R³ is a radical of formula (iii)

^R5-V o V

(iii)

6. A compound as claimed in any one of claims 1 to 4 wherein R³ is a radical of formula (iv)

(iv)

7. A compound as claimed in any one of claims 1 to 6 wherein R¹ and R² are both a radical of formul

(i)

8. A compound as claimed in any one of claims 1 to 6 wherein R¹ and R² are both a radical of formula (ii)

9. A compound as claimed in any one of claims 1 to 8 wherein R⁵

10. A compound as claimed in any one of claims 1 to 3, 5 to 6 or claim 9 wherein Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)_tCH2; and t is an integer from 1 to 10.

11. A compound as claimed in any one of claims 1 to 6 or claim 9 wherein Y is C; R¹ and R² are both H; A, E, B and D are CH₂ and X is O.

12. A compound as claimed in any one of claims 1 to 6 or claim 9 wherein Y is C; A is (CH₂)_U;

R¹ and R² are both H; B, X, D and E are all absent; and u is an integer from 1 to 10.

13. A compound as claimed in any one of claims 1 to 6 or claim 9 wherein:

Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P;

R¹ is H, NHZ or C_1-6alkyl;

R² is a radical of formula (i) or a radical of formula (ii)

Z is H, acyl, C(0)(CH₂)_wNHG, ^*CH₃ ^*C(0)- where ^*C denotes ¹³C or ¹⁴C, 4- carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6- sulfocoumarin-3-acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3- {[4-(2-cyclooctyn-1-ylmethyl)benzoyl]amino}propyl}-4-{2-[4- (dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate;

w is an integer from 1 to 11 ;

G is H, acyl, t-butoxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 9- fluorenylmethoxycarbonyl, benzyloxycarbonyl, *CH₃ ^*CO- where ^*C denotes ¹³C or ⁴C, 4-carboxytetramethylrhodamine, resorcinolphthalein, 7-amino-4-methyl-6- sulfocoumarin-3-acetic acid, 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene or 1-{3- {[4-(2-cyclooctyn-1-ylmethyl)benzoyl]amino}propyl}-4-{2-[4- (dimethylamino)phenyl]ethenyl}pyridinium hexafluorophosphate.

14. A compound as claimed in any one of claims 1 to 7 or claim 9 wherein Y is C; X is O; A, E and D ¹ and R², both the same, are a radical of formula (i)

(i)

R³ is a radical of formula (iii) o

; and and p is 1.

15. A compound as claimed in any one of claims 1 to 6 or claim 8 or claim 9 wherein Y is C; X is O; A, E and D are all CH₂; B is (CH₂)_P; R¹ and R², both the same, are a radical of formul

(ii)

and R³ is a radical of formula (iv)

(iv)

and p is 1.

16. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 11 wherein Y is C;

R¹ and ² are both H; A, E, B and D are CH₂; X is O; and R³ is a radical of formula (iii)

(iii)

17. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 11 wherein Y is C;

R¹ and R² are both H; A, E, B and D are CH₂; X is O; and R³ is a radical of formula (iv)

18. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 13 wherein Y is C;

X is O; (CH₂)_P; R¹ is H; R² is a radical of formula (i)

(i)

R³ is a radical of formula (iii)

(iii)

and p is 1.

A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 13 wherein Y is C; X is O; R¹ is H; R² is a radical of formula (ii)

(ii)

R³ is a radical of formula (iv)

(iv)

and p is 1.

20. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 13 wherein Y is C;

X is O; A, E and D are all CH₂; B is (CH₂)_P; R is H; R² is a radical of formula (ii)

(ii)

R³ is a radical of formula (iii)

R⁵-C

O

(iii) and p is 1.

A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 13 wherein Y is C; X is O; (CH₂)_P; R¹ is H; R² is a radical of formula (i)

0)

and R³ is a radical of formula (iv)

and p is 1. 22. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 12 wherein Y is C;

A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; u is an integer from 1 to 10; an ³ is a radical of formula (iii)

(iii) 23. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 12 wherein Y is C;

A is (CH₂)_U; R¹ and R² are both H; B, X, D and E are all absent; u is an integer from 1 to 10; an ³ is a radical of formula (iv)

(iv) 24. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 10 wherein Y is O;

B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)_tCH₂; t is an integer from 1 to 10; and R³ is a radical of formula (iii)

(iii)

5. A compound as claimed in any one of claims 1 to 6 or claim 9 or claim 10 wherein Y is O; B is O; R¹ and R² are absent; and either A, E, D and X are all CH₂ or A, D and X are all CH₂ and E is (CH₂CH₂0)_tCI-l₂; t is an integer from 1 to 10; and R³ is a radical of formula (iv)

(iv)

A compound as claimed in claim 1 , selected from the group consisting of:

59



61

63

64

(CH₂)₇ o _(CH_{2) 0} O O ^H O O (CH₂)

27. A compound selected from the group consisting of: