PH20 polypeptide variants, formulations and uses thereof

A modified PH20 polypeptide, comprising one or more amino acid replacements in an unmodified PH20 polypeptide, wherein:

• the unmodified PH20 polypeptide consists of the amino acid sequence selected from the group consisting of SEQ ID NOS:
• 3, 7 and 32-66;
• the modified PH20 polypeptide exhibits increased hyaluronidase activity that is at least 120% of the hyaluronidase activity compared to the unmodified PH20 polypeptide not containing the amino acid replacements);
• wherein:
• the activity of the unmodified PH20 polypeptide not containing the replacement(s) is 100%;
• hyaluronidase activity of the modified PH20 polypeptide and hyaluronidase activity of the unmodified PH20 polypeptide are measured under identical conditions:
• at least one amino acid replacement confers the increased hyaluronidase activity, and is selected from E at a position corresponding to position 12;
• E at a position corresponding to position 26;
• K at a position corresponding to position 26;
• R at a position corresponding to position 46;
• Q at a position corresponding to position 52;
• Q at a position corresponding to position 58;
• H at a position corresponding to position 68;
• Q at a position corresponding to position 68;
• A at a position corresponding to position 70;
• C at a position corresponding to position 70;
• F at a position corresponding to position 70;
• G at a position corresponding to position 70;
• H at a position corresponding to position 70;
• K at a position corresponding to position 70;
• L at a position corresponding to position 70;
• N at a position corresponding to position 70;
• P at a position corresponding to position 70;
• R at a position corresponding to position 70;
• S at a position corresponding to position 70;
• T at a position corresponding to position 70;
• A at a position corresponding to position 74;
• C at a position corresponding to position 74;
• G at a position corresponding to position 74;
• N at a position corresponding to position 74;
• P at a position corresponding to position 74;
• R at a position corresponding to position 74;
• S at a position corresponding to position 74;
• V at a position corresponding to position 74;
• W at a position corresponding to position 74;
• G at a position corresponding to position 84;
• R at a position corresponding to position 84;
• G at a position corresponding to position 97;
• H at a position corresponding to position 127;
• N at a position corresponding to position 127;
• G at a position corresponding to position 131;
• V at a position corresponding to position 131;
• R at a position corresponding to position 142;
• A at a position corresponding to position 166;
• F at a position corresponding to position 166;
• H at a position corresponding to position 166;
• L at a position corresponding to position 166;
• R at a position corresponding to position 166;
• Y at a position corresponding to position 166;
• K at a position corresponding to position 174;
• K at a position corresponding to position 206;
• M at a position corresponding to position 213;
• N at a position corresponding to position 213;
• E at a position corresponding to position 237;
• S at a position corresponding to position 238;
• N at a position corresponding to position 261;
• R at a position corresponding to position 277;
• T at a position corresponding to position 277;
• L at a position corresponding to position 309;
• N at a position corresponding to position 309;
• Q at a position corresponding to position 309;
• R at a position corresponding to position 309;
• T at a position corresponding to position 309;
• A at a position corresponding to position 310;
• G at a position corresponding to position 310;
• H at a position corresponding to position 315;
• I at a position corresponding to position 317;
• K at a position corresponding to position 317;
• R at a position corresponding to position 317;
• M at a position corresponding to position 318;
• G at a position corresponding to position 347;
• S at a position corresponding to position 347;
• M at a position corresponding to position 399;
• T at a position corresponding to position 399;
• W at a position corresponding to position 399;
• D at a position corresponding to position 407;
• Q at a position corresponding to position 407;
• F at a position corresponding to position 419;
• I at a position corresponding to position 419;
• R at a position corresponding to position 419;
• S at a position corresponding to position 419;
• K at a position corresponding to position 421;
• N at a position corresponding to position 421;
• Q at a position corresponding to position 421;
• R at a position corresponding to position 421;
• S at a position corresponding to position 421;
• A at a position corresponding to position 431;
• H at a position corresponding to position 431;
• K at a position corresponding to position 431;
• Q at a position corresponding to position 431;
• R at a position corresponding to position 431;
• S at a position corresponding to position 431;
• V at a position corresponding to position 431;
• L at a position corresponding to position 433;
• R at a position corresponding to position 433;
• V at a position corresponding to position 439;
• H at a position corresponding to position 440;
• R at a position corresponding to position 440;
• A at a position corresponding to position 443;
• M at a position corresponding to position 443;
• M at a position corresponding to position 445, and/or P at a position corresponding to position 445, with reference to amino acid positions set forth in SEQ ID NO:3;
• and
• the modified PH20 polypeptide has at least 95% sequence identity to the amino acid sequence selected from the group consisting of SEQ ID NOs:
• 3, 7, and 32-66.