Polypeptides

Claim 2

A derivative as claimed in claim 1 having at least one further modification selected from:- a) Leu¹⁵ of the native sequence replaced by a Glu¹⁵ residue; b) Lys²³ of the native sequence replaced by an Arg²³ residue; c) Gly²⁶ of the native sequence replaced by an Ala²⁶ residue; d) Gly²⁸ of the native sequence replaced by an Ala²⁸ residue; e) Ala 3O of the native sequence replaced by a Lys 3O or Arg 3O residue; f) Lys³⁴ of the native sequence replaced by an Arg³⁴ residue; g) Lys 4O of the native sequence replaced by an Arg 4O residue; h) Pro⁴⁴ of the native sequence replaced by an Ala⁴⁴ residue; i) Leu⁴⁹ of the native sequence replaced by a Lys⁴⁹ residue; j) Gly⁵¹ of the native sequence replaced by an Ala⁵¹ residue; k) Gly⁵⁵ of the native sequence replaced by an Ala⁵⁵ residue; l) Trp⁵⁸ of the native sequence replaced by a Lys⁵⁸ residue; m) Pro 6O of the native sequence replaced by a Ser 6O residue; n) Pro⁶⁵ of the native sequence replaced by a Ser⁶⁵ residue; o) Pro¹¹¹ of the native sequence replaced by a Glu¹¹¹ residue; p) Thr¹¹⁵ of the native sequence replaced by a Ser¹¹⁵ residue; q) Thr¹¹⁶ of the native sequence replaced by a Ser¹¹⁶ residue; and r) Tyr¹⁶⁵ of the native sequence replaced by an Arg¹⁶⁵ residue.

Claim 3

A derivative as claimed in claim 1 or claim 2 wherein the further modification comprises at least one of the following:- a) Gln¹¹, Pro 6O,65 of the native sequence replaced by Arg¹¹, Ser 6O,65 ; b) Ala¹¹¹, Thr 115,116 of the native sequence replaced by Glu¹¹¹, Ser 115,116 ; c) Gln 11, Trp⁵⁸, Tyr¹⁵⁸ of the native sequence replaced by Arg 11,165 , Lys⁵⁸; d) Leu¹⁵, Gly 26,28 , Ala 3O of the native sequence replaced by Glu¹⁵, Ala 26,28 , Lys 3O ;or e) Pro⁴⁴, Leu⁴⁹, Gly 51,55 , Trp⁵⁸ of the native sequence replaced by Lys 49,58 , Ala 44,51,55 . f) Leu¹⁵,Gly 26,28 ,Ala 3O of the native sequence replaced by Glu¹⁵, Ala 26,28 , Arg 3O ; or g) Pro⁶⁵ of the native sequence replaced by Ser⁶⁵.

Claim 4

A derivative as claimed in any one of the preceeding claims selected from:- [Arg¹¹,Ser 17,27,6O,65 ]hu G-CSF; [Glu¹⁵,Ser 17,27 ,Ala 26,28 ,Lys 3O ]hu G-CSF; [Arg¹¹,Glu¹⁵,Ser 17,27,6O,65 ,Ala 26,28 ,Lys 3O ]hu G-CSF; [Arg 11,165 Glu¹⁵ Ser 17,27,6O,65 Ala 26,28 ,Lys 3O,58 ]hu G-CSF; [Arg 11,23 ,Ser 17,27,6O,65 ]hu G-CSF; [Arg 11,4O ,Ser 17,27,6O,65 ]hu G-CSF; [Glu 15,111 ,Ser 17,27,6O,65,115,116 ,Ala 26,28 Lys 3O ]hu G-CSF [Ala¹,Thr³,Tyr⁴,Arg 5,11 ,Ser 17,27,6O,65 ]hu G-CSF [Glu¹⁵,Ser 17,27 ,Ala 26,28 ,Arg 3O ]hu G-CSF [Ser 17,27,6O,65 ]hu G-CSF [Arg¹¹,Ser 17,27,65 ]hu G-CSF [Ser 17,27,65 ]hu G-CSF said derivative, if desired having a presequence methionine.

Claim 5

A DNA sequence encoding all or part of the amino acid sequence of a derivative as claimed in any one of the preceding claims.

Claim 6

A recombinant vector containing a DNA sequence as defined in claim 5.

Claim 7

A process for the preparation of a recombinant vector as defined in claim 6 which comprises inserting a DNA sequence as defined in claim 5 into a vector.

Claim 8

A procaryotic or eucaryotic host cell stably transformed or transfected with a recombinant vector as defined in claim 6.

Claim 9

A process for the preparation of a procaryotic or eucaryotic host cell as defined in claim 8 which comprises transforming or transfecting a procaryotic or eucaroytic cell with a recombinant vector as defined in claim 6 whereby to yield a stably transformed or transfected procaryotic or eucaryotic host.

Claim 10

A process for the preparation of a derivative of naturally occurring G-CSF as defined in any one of claims 1 to 4 which comprises culturing a procaryotic or eucaryotic host cell as defined in claim 8 whereby to obtain said derivative.

Claim 11

A pharmaceutical composition comprising as active ingredient at least one derivative of naturally occurring G-CSF as claimed in any one of claims 1 to 4 in association with a pharmaceutically aceeptable carrier or excipient.

Claim 14

A process for extracting a derivative as claimed in any one of claims 1 to 4 from an inclusion body thereof which comprises 1) suspending said inclusion body in a detergent, 2) oxidation, 3) removal of detergent and 4) maintaining the solution obtained following removal of detergent at an elevated temperature whereby to precipitate contaminating bacterial protein, product oligomers and/or degradation products, whilst retaining said derivative in solution in active form.

Claim 15

A process as claimed in claim 14 wherein the derivative to be extracted has a solution stability of at least 85% at 1Omg/ml.